Systems and methods for multi-analysis

ABSTRACT

Systems and methods are provided for sample processing. A device may be provided, capable of receiving the sample, and performing one or more of a sample preparation, sample assay, and detection step. The device may be capable of performing multiple assays. The device may comprise one or more modules that may be capable of performing one or more of a sample preparation, sample assay, and detection step. The device may be capable of performing the steps using a small volume of sample.

CROSS-REFERENCE

This application claims priority to U.S. Provisional Patent ApplicationNo. 61/766,113, filed Feb. 18, 2013 and U.S. Provisional PatentApplication No. 61/766,119, filed Feb. 18, 2013. Both of the foregoingapplications are incorporated herein by reference in their entirety forall purposes.

BACKGROUND OF THE INVENTION

The majority of clinical decisions are based on laboratory and healthtest data, yet the methods and infrastructure for collecting such dataseverely limit the quality and utility of the data itself. Almost allerrors in laboratory testing are associated with human or pre-analyticprocessing errors, and the testing process can take days to weeks tocomplete. Often times by the time a practicing physician gets the datato effectively treat a patient or determine the most appropriateintervention, he or she has generally already been forced to treat apatient empirically or prophylactically as the data was not available atthe time of the visit or patient triage. Earlier access to higherquality testing information at the time of patient triage enablesearlier interventions and better management of disease progression toimprove outcomes and lower the cost of care.

Existing systems and methods for clinical testing suffer major drawbacksfrom the perspectives of patients, medical care professionals,taxpayers, and insurance companies. Today, consumers can undergo certainspecialized tests at clinics or other specialized locations. If a testis to be conducted and the result of which is to be eventually relied onby a doctor, physical samples are transported to a location whichperforms the test on the samples. For example, these samples maycomprise blood from a venous draw and are typically collected from asubject at the specialized locations. Accessibility of these locationsand the venipuncture process in and of itself is a major barrier incompliance and frequency of testing. Availability for visiting a bloodcollection site, the fear of needles—especially in children and elderlypersons who, for example, often have rolling veins, and the difficultyassociated with drawing large amounts of blood drives people away fromgetting tested even when it is needed. Thus, the conventional samplingand testing approach is cumbersome and requires a significant amount oftime to provide test results. Such methods are not only hampered byscheduling difficulties and/or limited accessibility to collection sitesfor subjects to provide physical samples but also by the batchprocessing of samples in centralized laboratories and the associatedturn around time in running laboratory tests. As a result, the overallturn around time involved in getting to the collection site, acquiringthe sample, transporting the sample, testing the sample and reportingand delivering results becomes prohibitive and severely limits thetimely provision of the most informed care from a medical professional.This often results in treatment of symptoms as opposed to underlyingdisease conditions or mechanisms of disease progression.

In addition, traditional techniques are problematic for certaindiagnoses. Some tests may be critically time sensitive, but take days orweeks to complete. Over such a time, a disease can progress past thepoint of treatment. In some instances, follow-up tests are requiredafter initial results, which take additional time as the patient has toreturn to the specialized locations. This impairs a medicalprofessional's ability to provide effective care. Furthermore,conducting tests at only limited locations and/or infrequently reducesthe likelihood that a patient's status can be regularly monitored orthat the patient will be able to provide the samples quickly or asfrequently as needed. For certain diagnoses or conditions, thesedeficiencies inevitably cause inadequate medical responses to changingand deteriorating physiological conditions. Traditional systems andmethods also affect the integrity and quality of a clinical test due todegradation of a sample that often occurs while transporting such samplefrom the site of collection to the place where analysis of the sample isperformed. For example, analytes decay at a certain rate, and the timedelay for analysis can result in loss of sample integrity. Differentlaboratories also work with different quality standards which can resultin varying degrees of error. Additionally, preparation and analysis ofsamples by hand permits upfront human error to occur at various samplecollection sites and laboratories. These and other drawbacks inherent inthe conventional setup make it difficult to perform longitudinalanalyses, especially for chronic disease management, with high qualityand reliability

Furthermore, such conventional analytical techniques are often not costeffective. Excessive time lags in obtaining test results lead to delaysin diagnoses and treatments that can have a deleterious effect on apatient's health; as a disease progresses further, the patient thenneeds additional treatment and too often ends up unexpectedly seeingsome form of hospitalization. Payers, such as health insurance companiesand taxpayers contributing to governmental health programs, end uppaying more to treat problems that could have been averted with moreaccessible and faster clinical test results.

SUMMARY OF THE INVENTION

Being able to detect a disease or the onset of a disease in time tomanage and treat it is a capability deeply sought after by patients andproviders alike but one that has yet to be realized in the currenthealthcare system where detection too often coincides with fatalprognoses.

A need exists for improved systems and methods for sample collection,sample preparation, assay, and/or detection. A further need exists forsystems and devices that perform one or more of the sample collection,preparation, assay, or detection steps. Systems and methods are neededat the time and place in which care is provided for rapid, frequentand/or more accurate diagnoses, ongoing monitoring, and facilitation andguidance of treatment. Systems and methods disclosed herein meet thisand related needs.

In accordance with an aspect of the invention, a system may comprise: aplurality of modules mounted on a support structure, wherein anindividual module of said plurality of modules comprises a samplepreparation station, assay station, and/or detection station, whereinthe system is configured to perform (a) at least one sample preparationprocedure selected from the group consisting of sample processing,centrifugation, separation, and chemical processing, and (b) multipletypes of assays selected from the group consisting of immunoassay,nucleic acid assay, receptor-based assay, cytometric assay, colorimetricassay, enzymatic assay, electrophoretic assay, electrochemical assay,spectroscopic assay, chromatographic assay, microscopic assay,topographic assay, calorimetric assay, turbidimetric assay,agglutination assay, radioisotope assay, viscometric assay, coagulationassay, clotting time assay, protein synthesis assay, histological assay,culture assay, osmolarity assay, and combinations thereof; and whereinthe multiple types of assays are performed with the aid of isolated(including but not limited to fluidically) assay units contained withinthe system. In some embodiments, separation includes magneticseparation.

Additional aspects of the invention may be directed to a system,comprising: a plurality of modules mounted on a support structure,wherein an individual module of said plurality of modules comprises asample preparation station, assay station, and/or detection station,wherein the system is configured to perform (a) at least one samplepreparation procedure selected from the group consisting of sampleprocessing, centrifugation, separation, and chemical processing, and (b)one or more types of assays selected from the group consisting ofimmunoassay, nucleic acid assay, receptor-based assay, cytometric assay,colorimetric assay, enzymatic assay, electrophoretic assay,electrochemical assay, spectroscopic assay, chromatographic assay,microscopic assay, topographic assay, calorimetric assay, turbidmetricassay, agglutination assay, radioisotope assay, viscometric assay,coagulation assay, clotting time assay, protein synthesis assay,histological assay, culture assay, osmolarity assay, and combinationsthereof; and wherein the system is configured to process or assay asample having a volume less than or equal to 250 μl, and the system hasa coefficient of variation less than or equal to 15%. In someembodiments, separation includes magnetic separation.

A system may be provided in accordance with another aspect of theinvention, said system comprising: a preparation station configured toperform sample preparation; and an assay station configured to performmultiple types of assays selected from the group consisting ofimmunoassay, nucleic acid assay, receptor-based assay, cytometric assay,colorimetric assay, enzymatic assay, electrophoretic assay,electrochemical assay, spectroscopic assay, chromatographic assay,microscopic assay, topographic assay, calorimetric assay, turbidmetricassay, agglutination assay, radioisotope assay, viscometric assay,coagulation assay, clotting time assay, protein synthesis assay,histological assay, culture assay, osmolarity assay, and combinationsthereof; and wherein the system is configured to perform said samplepreparation and said multiple types of assays within 4 hours or less.

In some aspects of the invention a system may be provided, comprising: aplurality of modules mounted on a support structure, wherein anindividual module of said plurality of modules comprises a samplepreparation station, assay station, and/or detection station, whereinthe system is configured to (a) prepare a sample for at least onephysical or chemical assay; and (b) perform said at least one physicalor chemical assay, and wherein at least one individual module of saidplurality comprises a cytometry station configured to perform cytometryon said sample.

Additional aspects of the invention are directed to a system,comprising: a sample preparation station, assay station, and detectionstation; and a control unit having computer-executable commands forperforming a point-of-service service at a designated location with theaid of at least one of said sample preparation station, assay stationand detection station, wherein the sample preparation station includes asample collection unit configured to collect a biological sample, andwherein the system is configured to assay a biological sample at acoefficient of variation less than or equal to 15%.

In accordance with aspects of the invention a system may comprise: ahousing; and a plurality of modules within said housing, an individualmodule of said plurality of modules comprising at least one stationselected from the group consisting of a sample preparation station,assay station, and detection station, wherein the system comprises afluid handling system configured to transfer a sample or reagent vesselwithin said individual module or from said individual module to anothermodule within the housing of said system.

A plug-and-play system may be provided in accordance with additionalaspect of the invention. The system may comprise: a supporting structurehaving a mounting station configured to support a module among aplurality of modules, said module being (a) detachable from saidmounting station or interchangeable with at least other module of theplurality; (b) configured to perform without the aid of another modulein said system (i) at least one sample preparation procedure selectedfrom the group consisting of sample processing, centrifugation, magneticseparation, or (ii) at least one type of assay selected from the groupconsisting of immunoassay, nucleic acid assay, receptor-based assay,cytometric assay, colorimetric assay, enzymatic assay, electrophoreticassay, electrochemical assay, spectroscopic assay, chromatographicassay, microscopic assay, topographic assay, calorimetric assay,turbidmetric assay, agglutination assay, radioisotope assay, viscometricassay, coagulation assay, clotting time assay, protein synthesis assay,histological assay, culture assay, osmolarity assay, and combinationsthereof; and (c) configured to be in electrical, electro-magnetical oroptoelectronic communication with a controller, said controller beingconfigured to provide one or more instructions to said module orindividual modules of said plurality of modules to facilitateperformance of the at least one sample preparation procedure or the atleast one type of assay.

Another aspect of the invention may be directed to a system, comprising:a sample preparation station, assay station, and/or detection station;and a control unit having computer-executable commands configured toperform a point-of-service service at a designated location, wherein thesystem is configured to perform multiple types of assays selected fromthe group consisting of immunoassay, nucleic acid assay, receptor-basedassay, cytometric assay, colorimetric assay, enzymatic assay,electrophoretic assay, electrochemical assay, spectroscopic assay,chromatographic assay, microscopic assay, topographic assay,calorimetric assay, turbidmetric assay, agglutination assay,radioisotope assay, viscometric assay, coagulation assay, clotting timeassay, protein synthesis assay, histological assay, culture assay,osmolarity assay, and combinations thereof.

Also, aspects of the invention may include a system, comprising: aplurality of modules mounted on a support structure, wherein anindividual module of said plurality of modules comprises a samplepreparation station, assay station, and/or detection station, whereinthe system is configured to perform (a) at least one sample preparationprocedure selected from the group consisting of sample processing,centrifugation, magnetic separation, and (b) multiple types of assaysselected from the group consisting of immunoassay, nucleic acid assay,receptor-based assay, cytometric assay, colorimetric assay, enzymaticassay, electrophoretic assay, electrochemical assay, spectroscopicassay, chromatographic assay, microscopic assay, topographic assay,calorimetric assay, turbidmetric assay, agglutination assay,radioisotope assay, viscometric assay, coagulation assay, clotting timeassay, protein synthesis assay, histological assay, culture assay,osmolarity assay, and combinations thereof; and wherein the multipletypes of assays are performed with the aid of three or more assay unitscontained within the system.

A system may be provided in accordance with another aspect of thesystem, said system comprising: a plurality of modules mounted on asupport structure, wherein an individual module of said plurality ofmodules comprises a sample preparation station, assay station, and/ordetection station, wherein the system is configured to perform (a) atleast one sample preparation procedure selected from the groupconsisting of sample processing, centrifugation, magnetic separation,and chemical processing, and (b) one or more types of assays selectedfrom the group consisting of immunoassay, nucleic acid assay,receptor-based assay, cytometric assay, colorimetric assay, enzymaticassay, electrophoretic assay, electrochemical assay, spectroscopicassay, chromatographic assay, microscopic assay, topographic assay,calorimetric assay, turbidmetric assay, agglutination assay,radioisotope assay, viscometric assay, coagulation assay, clotting timeassay, protein synthesis assay, histological assay, culture assay,osmolarity assay, and combinations thereof; and wherein the system isconfigured to process or assay a sample having a volume less than orequal to 250 μl, and the system has a coefficient of variation less thanor equal to 10%.

Furthermore, aspects of the invention may be directed to a system,comprising: an assay station configured to perform at least one type ofassay selected from the group consisting of immunoassay, nucleic acidassay, receptor-based assay, cytometric assay, colorimetric assay,enzymatic assay, electrophoretic assay, electrochemical assay,spectroscopic assay, chromatographic assay, microscopic assay,topographic assay, calorimetric assay, turbidmetric assay, agglutinationassay, radioisotope assay, viscometric assay, coagulation assay,clotting time assay, protein synthesis assay, histological assay,culture assay, osmolarity assay, and combinations thereof; and wherein acoefficient of variation of the at least one type of assay is less thanor equal to 10% when performed with said system.

In accordance with additional aspects of the invention, a system maycomprise: an assay station configured to perform multiple types ofassays selected from the group consisting of immunoassay, nucleic acidassay, receptor-based assay, cytometric assay, colorimetric assay,enzymatic assay, electrophoretic assay, electrochemical assay,spectroscopic assay, chromatographic assay, microscopic assay,topographic assay, calorimetric assay, turbidmetric assay, agglutinationassay, radioisotope assay, viscometric assay, coagulation assay,clotting time assay, protein synthesis assay, histological assay,culture assay, osmolarity assay, and combinations thereof; and a controlunit having computer-executable commands to perform said multiple typesof assays, wherein the system is configured to assay a biological samplehaving a volume less than or equal to 250 μl.

A system may be provided in accordance with additional aspects of theinvention, said system comprising: a preparation station configured toperform sample preparation; and an assay station configured to performmultiple types of assays selected from the group consisting ofimmunoassay, nucleic acid assay, receptor-based assay, cytometric assay,colorimetric assay, enzymatic assay, electrophoretic assay,electrochemical assay, spectroscopic assay, chromatographic assay,microscopic assay, topographic assay, calorimetric assay, turbidmetricassay, agglutination assay, radioisotope assay, viscometric assay,coagulation assay, clotting time assay, protein synthesis assay,histological assay, culture assay, osmolarity assay, and combinationsthereof, wherein the system is configured to perform said samplepreparation and said multiple types of assays within 4 hours or less.

Additionally, aspects of the invention may be directed to a system,comprising: a plurality of modules mounted on a support structure,wherein an individual module of said plurality of modules comprises asample preparation station, assay station, and/or detection station,wherein the system is configured to perform (a) at least one samplepreparation procedure selected from the group consisting of sampleprocessing, centrifugation, magnetic separation, and chemicalprocessing, and (b) multiple types of assays selected from the groupconsisting of immunoassay, nucleic acid assay, receptor-based assay,cytometric assay, colorimetric assay, enzymatic assay, electrophoreticassay, electrochemical assay, spectroscopic assay, chromatographicassay, microscopic assay, topographic assay, calorimetric assay,turbidmetric assay, agglutination assay, radioisotope assay, viscometricassay, coagulation assay, clotting time assay, protein synthesis assay,histological assay, culture assay, osmolarity assay, and combinationsthereof; and wherein the system is configured to process or assay asample having a volume less than or equal to 250 μl, and wherein thesystem is configured to detect from said sample a plurality of analytes,the concentrations of said plurality of analytes varying from oneanother by more than one order of magnitude.

Another aspect of the system may provide a system, comprising: a samplepreparation station, assay station, and/or detection station; and acontrol system having computer-executable commands configured to performa point-of-service service at a designated location, wherein the systemis configured to perform multiple types of assays selected from thegroup consisting of immunoassay, nucleic acid assay, receptor-basedassay, cytometric assay, colorimetric assay, enzymatic assay,electrophoretic assay, electrochemical assay, spectroscopic assay,chromatographic assay, microscopic assay, topographic assay,calorimetric assay, turbidmetric assay, agglutination assay,radioisotope assay, viscometric assay, coagulation assay, clotting timeassay, protein synthesis assay, histological assay, culture assay,osmolarity assay, and combinations thereof.

In accordance with additional aspects of the invention, a system maycomprise: a plurality of modules mounted on a support structure, whereinan individual module of said plurality of modules comprises a samplepreparation station, assay station, and/or detection station; whereinthe system is configured to perform multiple types of assays selectedfrom the group consisting of immunoassay, nucleic acid assay,receptor-based assay, cytometric assay, colorimetric assay, enzymaticassay, electrophoretic assay, electrochemical assay, spectroscopicassay, chromatographic assay, microscopic assay, topographic assay,calorimetric assay, turbidmetric assay, agglutination assay,radioisotope assay, viscometric assay, coagulation assay, clotting timeassay, protein synthesis assay, histological assay, culture assay,osmolarity assay, and combinations thereof, wherein at least one of saidmultiple types of assays is cytometry or agglutination.

A system, in accordance with additional aspects of the invention, maycomprise: a plurality of modules mounted on a support structure, whereinan individual module of said plurality of modules comprises a samplepreparation station, assay station, and/or detection station; acytometry station configured to perform cytometry on one or moresamples, wherein the system is configured to perform at least one assayselected from the group consisting of immunoassay, nucleic acid assay,receptor-based assay, cytometric assay, colorimetric assay, enzymaticassay, electrophoretic assay, electrochemical assay, spectroscopicassay, chromatographic assay, microscopic assay, topographic assay,calorimetric assay, turbidmetric assay, agglutination assay,radioisotope assay, viscometric assay, coagulation assay, clotting timeassay, protein synthesis assay, histological assay, culture assay,osmolarity assay, and combinations thereof.

Another aspect of the invention may provide a system, comprising: asample preparation station, assay station, and detection station; and acontrol unit having computer-executable commands for performing apoint-of-service service at a designated location with the aid of atleast one of said sample preparation station, assay station anddetection station, wherein the sample preparation station includes asample collection unit configured to collect a biological sample, andwherein the system is configured to assay a biological sample at acoefficient of variation less than or equal to 10%.

In some aspects of the invention, a system may comprise: a plurality ofmodules mounted on a support structure, wherein an individual module ofsaid plurality of modules comprises a sample preparation station, assaystation, and/or detection station, wherein the system is configured toperform (a) at least one sample preparation procedure selected from thegroup consisting of sample processing, centrifugation, magneticseparation, and (b) at least one physical or chemical assay, and whereinthe system is configured to assay a biological sample having a volumeless than or equal to 250 μl.

A system provided in accordance with an aspect of the invention maycomprise: a plurality of modules mounted on a support structure, whereinan individual module of said plurality of modules comprises a samplepreparation station, assay station, and/or detection station, whereinthe system is configured to perform (a) multiple sample preparationprocedures selected from the group consisting of sample processing,centrifugation, magnetic separation, physical separation and chemicalseparation, and (b) at least one type of assay selected from the groupconsisting of immunoassay, nucleic acid assay, receptor-based assay,cytometric assay, colorimetric assay, enzymatic assay, electrophoreticassay, electrochemical assay, spectroscopic assay, chromatographicassay, microscopic assay, topographic assay, calorimetric assay,turbidmetric assay, agglutination assay, radioisotope assay, viscometricassay, coagulation assay, clotting time assay, protein synthesis assay,histological assay, culture assay, osmolarity assay, and combinationsthereof.

Furthermore, some aspects of the invention may provide a system,comprising: a housing; and a plurality of modules within said housing,an individual module of said plurality of modules comprising at leastone station selected from the group consisting of a sample preparationstation, assay station, and detection station, wherein the systemcomprises a fluid handling system configured to transfer a sample orreagent vessel within said individual module or from said individualmodule to another module within the housing of said system.

Systems above or elsewhere herein, alone or in combination, may comprisea fluid handling system, wherein said fluid handling system comprises apipette configured to uptake, dispense, and/or transfer said biologicalsample.

Systems above or elsewhere herein may comprise an imaging deviceconfigured to image one or more of the group consisting of thebiological sample collected, processing of the biological sample, andreaction performed on the systems above or elsewhere herein, alone or incombination. The imaging device may be a camera or a sensor that detectsand/or record electromagnetic radiation and associated spacial and/ortemporal dimensions.

Systems above or elsewhere herein, alone or in combination may beconfigured to detect from said sample a plurality of analytes, theconcentrations of said plurality of analytes varying from one another bymore than one order of magnitude.

A sample collection unit configured to draw a fluid or tissue samplefrom a subject may be provide in systems above or elsewhere herein,alone or in combination.

Systems above or elsewhere herein, alone or in combination may have acoefficient of variation less than or equal to 10%.

An automated method for processing a sample at a point-of-servicelocation may be provided, said method comprising: providing the sampleto systems above or elsewhere herein, alone or in combination; andallowing said system to process said sample to yield a detectable signalindicative of completion of said processing.

In practicing the method above or elsewhere herein, alone or incombination, the processing step may assess histology of the sample ormorphology of the sample. The processing step may assesses the presenceand/or concentration of an analyte in the sample in methods above orelsewhere herein, alone or in combination.

In systems above or elsewhere herein, alone or in combination, thesample preparation station may comprise a sample collection unitconfigured to collect a biological sample from a subject.

A supporting structure may be a housing that encloses the plurality ofmodules, said housing optionally provides a power source orcommunication unit, in systems above or elsewhere herein, alone or incombination.

The systems above or elsewhere herein, alone or in combination, maystore and/or transmit electronic data representative of the image to anexternal device via a communication unit comprised in the system.

In some embodiments, systems above or elsewhere herein, alone or incombination may further comprise a centrifuge.

Systems above or elsewhere herein, alone or in combination, may beconfigured to perform two-way communication with an external device viaa communication unit comprised in said system, wherein the communicationunit is configured to send data to said external device and receiveinstructions with said system.

A method of detecting presence or concentration of an analyte suspectedto be present in a biological sample from a subject may be provided,said method comprising: providing the biological sample to systems aboveor elsewhere herein, alone or in combination; and performing at leastone type of assay selected from the group consisting of immunoassay,nucleic acid assay, receptor-based assay, cytometric assay, colorimetricassay, enzymatic assay, electrophoretic assay, electrochemical assay,spectroscopic assay, chromatographic assay, microscopic assay,topographic assay, calorimetric assay, turbidmetric assay, agglutinationassay, radioisotope assay, viscometric assay, coagulation assay,clotting time assay, protein synthesis assay, histological assay,culture assay, osmolarity assay, and combinations thereof, to yield adetectable signal indicative of the presence or concentration of saidanalyte.

Methods above or elsewhere herein, alone or in combination, may furthercomprise the step of generating a report comprising information relatingto a time dependent change of the presence or concentration of saidanalyte.

Methods above or elsewhere herein, alone or in combination, may furthercomprise the step of generating a report comprising information relatingto diagnosis, prognosis and/or treatment of a medical condition for saidsubject based on a time dependent change of the presence orconcentration of said analyte.

In some situations, chemical processing is selected from the groupconsisting of heating and chromatography. In some embodiments,receptor-based assay includes protein assay. In some embodiments,systems provided herein, alone or in combination, are configured forautonomous operation.

In some embodiments, systems, alone or in combination, are configured todetect from a sample a plurality of analytes, the concentrations of saidplurality of analytes varying from one another by more than one order ofmagnitude. The concentrations of said plurality of analytes may varyfrom one another by more than two orders of magnitude. In some cases,the concentrations of said plurality of analytes may vary from oneanother by more than three orders of magnitude. The multiple types ofassays may be performed with the aid of four or more assay unitscontained within the system. In some situations, systems are configuredto draw a fluid or tissue sample from a subject. In an embodiments,systems are configured to draw a blood sample from a finger of thesubject

In some embodiments, a system, alone or in combination, has acoefficient of variation less than or equal to 5%. In other embodiments,a system, alone or in combination, has a coefficient of variation lessthan or equal to 3%. In other embodiments, a system, alone or incombination, has a coefficient of variation less than or equal to 2%.The coefficient of variation in some cases is determined according toσ/μ wherein ‘σ’ is the standard deviation and ‘μ’ is the mean acrosssample measurements.

In some situations, systems provided herein are configured to performmultiple types of assays selected from the group consisting ofimmunoassay, nucleic acid assay, receptor-based assay, cytometric assay,colorimetric assay, enzymatic assay, electrophoretic assay,electrochemical assay, spectroscopic assay, chromatographic assay,microscopic assay, topographic assay, calorimetric assay, turbidmetricassay, agglutination assay, radioisotope assay, viscometric assay,coagulation assay, clotting time assay, protein synthesis assay,histological assay, culture assay, osmolarity assay, and combinationsthereof.

In some situations, systems provided herein have an accuracy of plus orminus 5% across sample assays, or plus or minus 3% across sample assays,or plus or minus 1% across sample assays, or plus or minus 5% acrosssample assays, or plus or minus 3% across sample assays, or plus orminus 1% across sample assays. In some embodiments, the coefficient ofvariation of the at least one type of assay is less than or equal to 5%,or less than or equal to 3%, or less than or equal to 2%.

In some cases, a system may further comprise a plurality of modulesmounted on a support structure, wherein an individual module of saidplurality of modules comprises a sample preparation station, assaystation, and/or detection station. Said individual module may comprise asample preparation station, assay station and detection station. In somecases, a system further comprises a sample preparation station, assaystation and detection station.

In some embodiments, systems above or elsewhere herein, alone or incombination, are configured to perform at least one sample preparationprocedure selected from the group consisting of sample processing,centrifugation, magnetic separation and chemical processing. Thechemical processing may be selected from the group consisting of heatingand chromatography.

In some embodiments, systems above or elsewhere herein, alone or incombination, include computer-executable commands. Thecomputer-executable commands may be provided by a server incommunication with the system.

In some embodiments, systems above or elsewhere herein, alone or incombination, include least one sample preparation procedure selectedfrom the group consisting of sample processing, centrifugation, magneticseparation, and chemical processing. Such systems can be configured toassay a sample at a rate of at least 0.25 assays/hour, or at least 0.5assays/hour, or at least 1 assay/hour, or at least 2 assays/hour. Suchsystem may include a control unit having computer-executable commandsfor performing a point-of-service service at a designated location. Thecomputer-executable commands may be provided by a server incommunication with the system. In some embodiments, systems above orelsewhere herein, alone or in combination, are configured to assay asample and report a result to a remote system within a time period of atleast about 6 hours, or 5 hours, or 3 hours, or 2 hours, or 1 hour, or30 minutes, or 10 minutes, or 1 minute, or 30 seconds, or 10 seconds, or5 seconds, or 1 seconds, or 0.1 seconds. For such systems, theconcentrations of a plurality of analytes may vary from one another bymore than two orders of magnitude, or three orders of magnitude.

In some embodiments, systems above or elsewhere herein, alone or incombination, are configured to correlate the concentrations of analyteswith compliance or non-compliance with a medical treatment.

In some embodiments, a system above or elsewhere herein, alone or incombination, includes a sample preparation station one or more samplecollection units. The one or more sample collection units may include alancet and/or needle. The needle may include a microneedle. The one ormore sample collection units may be configured to collect a biologicalsample.

In some embodiments, a system above or elsewhere herein, alone or incombination, includes a sample preparation station, assay station anddetection station.

In some embodiments, a system above or elsewhere herein, alone or incombination, is configured to perform multiple types of assays with theaid of fluidically isolated assay units contained within the system. Insome cases, the multiple types of assays are performed on an unprocessedtissue sample. In an example, the unprocessed tissue sample includesunprocessed blood.

In some embodiments, a system above, alone or in combination, isconfigured to perform cytometry. In other embodiments, a system above,alone or in combination, is configured to perform agglutination andcytometry. In other embodiments, a system above, alone or incombination, is configured to perform agglutination, cytometry andimmunoassay.

In some embodiments, a system above, alone or in combination, isconfigured to assay a biological sample at a coefficient of variationless than or equal to 10%, or less than or equal to 5%, or less than orequal to 3%.

In some embodiments, a system above, alone or in combination, isconfigured to perform at least one physical or chemical assay, such ascytometry. In some cases, the at least one physical or chemical assayfurther includes agglutination. In some cases, the at least one physicalor chemical assay further includes immunoassay.

In some embodiments, a system above, alone or in combination, isconfigured to process or assay a biological sample having a volume lessthan or equal to 100 μl. In other embodiments, a system above, alone orin combination, is configured to process or assay a sample having avolume less than or equal to 50 μl. In other embodiments, a systemabove, alone or in combination, is configured to process or assay asample having a volume less than or equal to 1 μl. In other embodiments,a system above, alone or in combination, is configured to process orassay a sample having a volume less than or equal to 500 nanoliters(nL).

In some embodiments, a system above, alone or in combination, is a pointof service system

In some embodiments, a system above, alone or in combination, isconfigured to perform two or more types of assays selected from thegroup consisting of immunoassay, nucleic acid assay, receptor-basedassay, cytometric assay, colorimetric assay, enzymatic assay,electrophoretic assay, electrochemical assay, spectroscopic assay,chromatographic assay, microscopic assay, topographic assay,calorimetric assay, turbidmetric assay, agglutination assay,radioisotope assay, viscometric assay, coagulation assay, clotting timeassay, protein synthesis assay, histological assay, culture assay,osmolarity assay, and combinations thereof. In some cases, the system,alone or in combination with other systems, is configured to performthree or more types of assays selected from said group.

In some embodiments, a system above, alone or in combination, isconfigured to perform at least one type of assay with the aid offluidically isolated assay units contained within the system. In somecases, the fluidically isolated assay units are tips. In some cases,each of the tips has a volume of at most 250 microliters (μl, also “ul”herein), or at most 100 μl, or at most 50 μl, or at most 1 μl, or atmost 500 nanoliters (nl).

In some embodiments, an individual module of a plurality of modulescomprises a fluid uptake or retention system. In some cases, the fluiduptake and/or retention system is a pipette.

In some embodiments, a system above, alone or in combination, isconfigured for two-way communication with a point of service server.

In some embodiments, a system above, alone or in combination, has afluid handling system having a coefficient of variation less than orequal to 10%, or less than or equal to 5%, or less than or equal to 3%,or less than or equal to 10%, or less than or equal to 5%, or less thanor equal to 3%. In some embodiments, the fluid handling system includesan optical fiber.

In some embodiments, a fluid handling system includes a fluid uptakeand/or retention system. In some cases, a fluid handling system includesa pipette. In some embodiments, the fluid handling system is attached toeach individual module among a plurality of modules of a systemdescribed above, alone or in combination with other systems. In someembodiments, a system above, alone or in combination, includes a housingthat comprises a rack for supporting the plurality of modules. Thehousing can be dimensioned to be no more than 3 m³, or no more than 2m³.

In some embodiments, a system above, alone or in combination, comprisesa control system having programmable commands for performing apoint-of-service service at a designated location.

In some embodiments, a system above, alone or in combination, includes afluid handling system. In some cases, the fluid handling system includesa pipette selected from the group consisting of a positive displacementpipette, air displacement pipette and suction-type pipette.

In some embodiments, a system above, alone or in combination, includes aplurality of modules. In some cases, an individual module comprisesfluid handling tips configured to perform one or more of proceduresselected from the group consisting of centrifugation, sample separation,immunoassay, nucleic acid assay, receptor-based assay, cytometric assay,colorimetric assay, enzymatic assay, electrophoretic assay,electrochemical assay, spectroscopic assay, chromatographic assay,microscopic assay, topographic assay, calorimetric assay, turbidimetricassay, agglutination assay, radioisotope assay, viscometric assay,coagulation assay, clotting time assay, protein synthesis assay,histological assay, culture assay, osmolarity assay, and combinationsthereof. In some situations, the nucleic acid assay is selected from thegroup consisting of nucleic acid amplification, nucleic acidhybridization, and nucleic acid sequencing.

In some embodiments, a system above, alone or in combination, includes aplurality of modules, and each individual module of said plurality ofmodules comprises (a) a fluid handling system configured to transfer asample within said individual module or from said individual module toanother module within said system, (b) a plurality of assay unitsconfigured to perform multiple types of assays, and (c) a detectorconfigured to detect signals generated from said assays. In somesituations, the multiple types of assays are selected from the groupconsisting of immunoassay, nucleic acid assay, receptor-based assay,cytometric assay, colorimetric assay, enzymatic assay, electrophoreticassay, electrochemical assay, spectroscopic assay, chromatographicassay, microscopic assay, topographic assay, calorimetric assay,turbidimetric assay, agglutination assay, radioisotope assay,viscometric assay, coagulation assay, clotting time assay, proteinsynthesis assay, histological assay, culture assay, osmolarity assay,and combinations thereof.

In some embodiments, a system above, alone or in combination, includes aplurality of modules, and each individual module comprises a centrifuge.

In some embodiments, a system above, alone or in combination, furthercomprises a module providing a subset of the sample preparationprocedures or assays performed by at least one module of said system.

In some embodiments, a system above, alone or in combination, comprisesan assay station that includes a thermal block.

In some embodiments, a sample includes at least one material selectedfrom the group consisting of fluid sample, tissue sample, environmentalsample, chemical sample, biological sample, biochemical sample, foodsample, or drug sample. In some cases, the sample includes blood orother bodily fluid, or tissue.

In some embodiments, a system above, alone or in combination, isconfigured for two-way communication with a point of service server. Insome cases, the two-way communication is wireless.

In some embodiments, a system above, alone or in combination, includes aplurality of modules, and each member of the plurality of modules isswappable with another module.

In some embodiments, a system above, alone or in combination, includesan assay station that comprises discrete assay units. In some cases, thediscrete assay units are fluidically isolated assay units.

In some embodiments, a system above, alone or in combination, isconfigured for longitudinal analysis at a coefficient of variation lessthan or equal to 10%, or less than or equal to 5%, or less than or equalto 3%.

In some embodiments, a system above, alone or in combination, includes afluid handling system that includes an optical fiber.

In some embodiments, a system above, alone or in combination, includes afluid handling system that includes a pipette.

In some embodiments, a system above, alone or in combination, comprisesan image analyzer.

In some embodiments, a system above, alone in combination, comprises atleast one camera in a housing of the system. In some cases, the at leastone camera is a charge-coupled device (CCD) camera. In some situations,the at least one camera is a lens-less camera.

In some embodiments, a system above, alone or in combination, comprisesa controller that includes programmable commands for performing apoint-of-service service at a designated location.

In some embodiments, a system above, alone or in combination, is aplug-and-play system configured to provide a point-of-service service.In some cases, the point-of-service service is a point of care serviceprovided to a subject having a prescription from the subject'scaretaker, said prescription being prescribed for testing the presenceor concentration of an analyte from said subject's biological sample.

In some embodiments, a system above, alone or in combination, includes aplurality of modules, and each member of the plurality of modulescomprises a communication bus in communication with a station configuredto perform the at least one sample preparation procedure or the at leastone type of assay.

In some embodiments, a system above, alone or in combination, includes asupporting structure. In some cases, the supporting structure is a rack.In some situations, the rack does not include a power or communicationcable; in other situations, the rack includes a power or communicationcable. In some embodiments, the supporting (or support) structureincludes one or more mounting stations. In some cases, the supportingstructure includes a bus in communication with a mounting station ofsaid one or more mounting stations.

In some embodiments, the bus is for providing power to individualmodules of the system. In some embodiments, the bus is for enablingcommunication between a controller of the system (e.g., plug-and-playsystem) and individual modules of the system. In some situations, thebus is for enabling communication between a plurality of modules of thesystem, or for enabling communication between a plurality of modules ofa plurality of systems.

In some embodiments, a system, alone or in combination, includes aplurality of modules, and each individual modules of the plurality ofmodules is in wireless communication with a controller of the system. Insome cases, wireless communication is selected from the group consistingof Bluetooth communication, radiofrequency (RF) communication andwireless network communication.

In some embodiments, a method for processing a sample, alone or incombination with other methods, comprises providing a system above,alone or in combination. The system comprises multiple modulesconfigured to perform simultaneously (a) at least one sample preparationprocedure selected from the group consisting of sample processing,centrifugation, magnetic separation and chemical processing, and/or (b)at least one type of assay selected from the group consisting ofimmunoassay, nucleic acid assay, receptor-based assay, cytometric assay,colorimetric assay, enzymatic assay, electrophoretic assay,electrochemical assay, spectroscopic assay, chromatographic assay,microscopic assay, topographic assay, calorimetric assay, turbidimetricassay, agglutination assay, radioisotope assay, viscometric assay,coagulation assay, clotting time assay, protein synthesis assay,histological assay, culture assay, osmolarity assay, and combinationsthereof within a module. Next, the system (or a controller of thesystem) tests for the unavailability of resources or the presence of amalfunction of (a) the at least one sample preparation procedure or (b)the at least one type of assay. Upon detection of the malfunction withinat least one module, the system uses another module within the system oranother system in communication with the system to perform the at leastone sample preparation procedure or the at least one type of assay.

In some cases, the system processes the sample at a point of servicelocation.

In some cases, the system is in wireless communication with anothersystem.

In some cases, multiple modules of the system are in electrical,electro-magnetic or optoelectronic communication with one another.

In some cases, multiple modules of the system are in wirelesscommunication with one another.

An aspect of the invention includes a fluid handling apparatuscomprising: a plurality of pipette heads, wherein an individual pipettehead comprises a pipette nozzle configured to connect with a tip that isremovable from the pipette nozzle; a plurality of plungers that areindividually movable, wherein at least one plunger is within a pipettehead and is movable within the pipette head; and a motor configured toeffect independent movement of individual plungers of the plurality.

Another aspect of the invention includes a fluid handling apparatuscomprising a plurality of pipette heads, wherein an individual pipettehead comprises a pipette nozzle configured to connect with a tip that isremovable from the pipette nozzle; a plurality of plungers that areindividually movable, wherein at least one plunger is within a pipettehead and is movable within the pipette head; and an actuator configuredto effect independent movement of individual plungers of the plurality.

Another aspect of the invention includes a fluid handling apparatuscomprising a plurality of pipette heads, wherein an individual pipettehead comprises a pipette nozzle configured to connect with a tip that isremovable from said pipette nozzle, wherein the fluid handling apparatusis capable of dispensing and/or aspirating 0.5 microliters (“uL”) to 5milliliters (“mL”) of fluid while functioning with a coefficient ofvariation of 5% or less.

A fluid handling apparatus may be provided in accordance with an aspectof the invention, the apparatus comprising: at least one pipette head,wherein an individual pipette head comprises a pipette nozzle configuredto connect with a tip that is removable from said nozzle; at least oneplunger within a pipette head of said plurality, wherein the plunger isconfigured to be movable within the pipette head; and at least one motorconfigured to permit movement of the plurality of plunger that is notsubstantially parallel to the removable tip.

Another aspect of the invention provides a fluid handling apparatuscomprising at least one pipette head, wherein an individual pipette headcomprises a pipette nozzle configured to connect with a tip that isremovable from said nozzle; at least one plunger within a pipette headof said plurality, and wherein the plunger is configured to be movablewithin the pipette head; and at least one actuator configured to permitmovement of the plurality of plungers that are not substantiallyparallel to the removable tip.

Another aspect of the invention may provide a fluid handling apparatuscomprising: at least one pipette head, wherein an individual pipettehead comprises a pipette nozzle configured to connect with a tip that isremovable from said nozzle, wherein said at least one pipette head has afluid path of a given length that terminates at the pipette nozzle, andwherein the length of the fluid path is adjustable without affectingmovement of fluid from the tip when the tip and the pipette nozzle areengaged.

Another aspect of the invention provides a fluid handling apparatuscomprising at least one pipette head, wherein an individual pipette headcomprises a pipette nozzle configured to connect with a tip that isremovable from said nozzle, wherein said at least one pipette head has afluid path of a given length that terminates at the pipette nozzle, andwherein the length of the fluid path is adjustable without affectingmovement of fluid from the tip when the tip and the pipette nozzle areengaged.

Additionally, aspects of the invention may include a fluid handlingapparatus comprising: a removable tip; and at least one pipette head,wherein an individual pipette head comprises a pipette nozzle configuredto connect with the tip that is removable from said pipette nozzle,wherein the apparatus is operably connected to an image capture devicethat is configured to capture an image within and/or through the tip.

An aspect of the invention may be directed to a sample processingapparatus comprising: a sample preparation station, assay station,and/or detection station; a control unit having computer-executablecommands for performing a point-of-service service at a designatedlocation with the aid of at least one of said sample preparationstation, assay station and detection station; and at least one pipettehaving a pipette nozzle configured to connect with a tip that isremovable from said pipette nozzle, wherein said pipette is configuredto transport a fluid no more than 250 uL within or amongst saidpreparation station, assay station and/or detection station.

A fluid handling apparatus may be provided in accordance with anadditional aspect of the invention. The fluid handling apparatus maycomprise: a plurality of pipette heads, wherein an individual pipettehead comprises a pipette nozzle configured to connect with a tip that isremovable from said pipette nozzle, wherein the fluid handling apparatusis capable of dispensing and/or aspirating 1 uL to 5 mL of fluid whilefunctioning with a coefficient of variation of 4% or less.

In accordance with another aspect of the invention, a fluid handlingapparatus may comprise: at least one pipette head operably connected toa base, wherein an individual pipette head comprises a pipette nozzleconfigured to connect with a removable tip; and at least one plungerwithin a pipette head of said plurality, wherein the plunger isconfigured to be movable within the pipette head, wherein the pipettenozzle is movable relative to the base, such that the pipette nozzle iscapable of having (a) a retracted position, and (b) an extended positionwherein the pipette nozzle is further away from the base than in theretracted position.

Also, an aspect of the invention may be directed to a fluid handlingapparatus comprising: a supporting body, extending therefrom a pluralityof pipette heads comprising a positive displacement pipette head,comprising a positive displacement pipette nozzle configured to connectwith a first removable tip; and an air displacement pipette head,comprising an air displacement pipette nozzle configured to connect toan air displacement pipette tip.

An aspect of the invention may be directed to a fluid handling apparatuscomprising: a plurality of pipette heads, wherein an individual pipettehead comprises a pipette nozzle configured to connect with a removabletip; a plurality of plungers, wherein at least one plunger is within apipette head of said plurality, and is configured to be movable withinthe pipette head, and said plurality of plungers are independentlymovable; and a motor configured to permit independent movement of theplurality of plungers.

Additional aspects of the invention may provide a fluid handlingapparatus comprising: a plurality of pipette heads, wherein anindividual pipette head comprises a pipette nozzle configured to connectwith a removable tip; a plurality of tip removal mechanisms, wherein atleast one tip removal mechanism is configured to be movable with respectto the pipette nozzle and to remove an individually selected tip fromthe pipette nozzle, and said plurality of tip removal mechanisms areindependently movable; and a motor configured to permit independentmovement of the plurality of tip removal mechanisms.

A fluid handling apparatus may be provided in accordance with anotheraspect of the invention, said apparatus comprising: a plurality ofpipette heads, wherein an individual pipette head comprises a pipettenozzle configured to connect with a removable tip, wherein the fluidhandling apparatus has a height, width, and length each of whichdimension does not exceed 20 cm.

Aspects of the invention may be directed to a fluid handling apparatuscomprising: a plurality of pipette heads, wherein an individual pipettehead comprises a pipette nozzle configured to connect with a removabletip, wherein the fluid handling apparatus is capable of dispensingand/or aspirating 1 uL to 3 mL of fluid while functioning with acoefficient of variation of 5% or less.

Additionally, a fluid handling apparatus may comprise: at least onepipette head, wherein an individual pipette head comprises a pipettenozzle configured to connect with a removable tip; and at least onemotor comprising a rotor and a stator, wherein the rotor is configuredto rotate about an axis of rotation, wherein the axis of rotation issubstantially perpendicular to the removable tip, accordance with anaspect of the invention.

Another aspect of the invention may be directed to a fluid handlingapparatus comprising: at least one pipette head, wherein an individualpipette head comprises a pipette nozzle configured to connect with aremovable tip; at least one plunger within a pipette head of saidplurality, wherein the plunger is configured to be movable within thepipette head; and at least one motor configured to permit movement ofthe plurality of plunger that is not substantially parallel to theremovable tip.

In accordance with additional aspects of the invention, a fluid handlingapparatus may comprise: at least one pipette head, wherein an individualpipette head comprises a pipette nozzle configured to connect with aremovable tip; and at least one plunger within a pipette head of saidplurality, wherein the plunger is configured to be movable within thepipette head, and wherein the plunger comprises a first section and asecond section wherein at least a portion of the first section isconfigured to slide relative to the second section, thereby permittingthe plunger to extend and/or collapse.

Another aspect of the invention may be directed to a fluid handlingapparatus comprising: at least one pipette head, wherein an individualpipette head comprises a pipette nozzle configured to connect with aremovable tip, wherein said at least one pipette head has a fluid pathof a given length that terminates at the pipette nozzle, and wherein thelength of the fluid path is adjustable without affecting movement offluid from the tip when the tip and the pipette nozzle are engaged.

A fluid handling apparatus, in accordance with an aspect of theinvention, may comprise: at least one pipette head operably connected toa base, wherein an individual pipette head comprises a pipette nozzleconfigured to connect with a removable tip; and at least one plungerwithin a pipette head of said plurality, wherein the plunger isconfigured to be movable within the pipette head, wherein the pipettenozzle is movable relative to the base, such that the pipette nozzle iscapable of having (a) a retracted position, and (b) an extended positionwherein the pipette nozzle is further away from the base than in theretracted position.

Furthermore, aspects of the invention may be directed to a method offluid handling comprising: providing at least one pipette head operablyconnected to a base, wherein an individual pipette head comprises apipette nozzle configured to connect with a removable tip; providing atleast one plunger within a pipette head of said plurality, wherein theplunger is configured to be movable within the pipette head; andretracting the pipette nozzle relative to the base in first directionprior to and/or concurrently with translating the pipette head in asecond direction substantially non-parallel to the first direction.

Another aspect of the invention may provide a method of fluid handlingcomprising: providing at least one pipette head operably connected to abase, wherein an individual pipette head comprises a pipette nozzleconfigured to connect with a removable tip; retracting and/or extendingthe pipette nozzle relative to the base; and dispensing and/oraspirating a fluid with the tip during said retracting and/or extending.

In accordance with some aspects of the invention, a fluid handlingapparatus may comprise: a supporting body, extending therefrom aplurality of pipette heads comprising a first pipette head of saidplurality, comprising a first pipette nozzle configured to connect witha first removable tip; a second pipette head of said plurality,comprising a second pipette nozzle configured to connect to a secondremovable tip; wherein the first removable tip is configured to hold upto a first volume of fluid, and the second removable tip is configuredto hold up to a second volume of fluid, wherein the first volume isabout 250 microliters, and the second volume is about 2 mL.

Aspects of the invention may be directed to a fluid handling apparatuscomprising: a supporting body, extending therefrom a plurality ofpipette heads comprising a positive displacement pipette head,comprising a positive displacement pipette nozzle configured to connectwith a first removable tip; and an air displacement pipette head,comprising an air displacement pipette nozzle configured to connect toan air displacement pipette tip.

Another aspect of the invention may provide a method of transportingcomponents within a device comprising: providing a plurality of pipetteheads, wherein an individual pipette head comprises a pipette nozzleconfigured to connect with a removable tip, wherein the individualpipette head is capable of dispensing and/or aspirating a fluid with thetip; engaging a sample processing component using at least one pipettehead of said plurality; and transporting the sample processing componentusing at least one pipette head of said plurality.

A fluid handling apparatus may be provided in accordance with anotheraspect of the invention, comprising: a removable tip; and at least onepipette head, wherein an individual pipette head comprises a pipettenozzle configured to connect with the removable tip, wherein theapparatus is operably connected to a light source that provides lightinto the tip.

Additionally, aspects of the invention may be directed to a fluidhandling apparatus comprising: a removable tip; and at least one pipettehead, wherein an individual pipette head comprises a pipette nozzleconfigured to connect with the removable tip, wherein the apparatus isoperably connected to an image capture device that is configured tocapture an image within and/or through the tip.

In accordance with an aspect of the invention, a fluid handlingapparatus may comprise: a removable tip; at least one pipette head,wherein an individual pipette head comprises a pipette nozzle configuredto connect with the removable tip; and a processor operably connected tothe removable tip and/or the at least one pipette head, wherein theapparatus is configured to vary and/or maintain the position of theremovable tip based on instructions from the processor.

A fluid handling apparatus comprising: a movable support structure; aplurality of pipette heads sharing the movable support structure,wherein an individual pipette head comprises a pipette nozzle configuredto connect with a removable tip, wherein the plurality of pipette headsare less than or equal to 4 mm apart from center to center, may beprovided in accordance with an aspect of the invention.

In some embodiments, a fluid handling apparatus above, alone or incombination with other systems, operates with a coefficient of variationless than or equal to about 10%. In some cases, the fluid handlingapparatus is capable of metering a fluid volume of 50 uL or less

In some embodiments, a system above, alone or in combination, includesone or more pipettes having pipette nozzles that are flexibly movable ina direction. In some cases, the pipette nozzles are spring-loaded.

In some embodiments, a system above, alone or in combination, hasremovable tips that are pipette tips having an interior surface, andexterior surface, and an open end.

In some embodiments, a system above, alone or in combination, has asolenoid for each plunger to determine whether individual plungers areto be moved.

In some embodiments, a system above, alone or in combination, has anactuator (or an actuation mechanism). The actuator in some casesincludes a motor. The motor may cause actuation of selected actuationmechanisms.

In some embodiments, a system above, alone or in combination, has afluid handling apparatus. The fluid handling apparatus may be configuredto aspirate or dispense no more than 250 uL at an individual fluidorifice. The fluid handling apparatus may be configured to aspirateand/or dispense a fluid that was collected from a subject via afingerstick. In some situations, the fingerstick is on a point ofservice device.

In some embodiments, a system above, alone or in combination, has aplurality of plungers that are capable of removing at least oneindividually selected tip from the pipette nozzle.

In some embodiments, a system above, alone or in combination, comprisesa plurality of external actuation mechanisms that external to a pipettehead of the system, wherein the plurality of external actuationmechanisms are capable of removing at least one individually selectedtip from the pipette nozzle. In some situations, an additional motorpermits independent movement of the plurality of external actuationmechanisms. In some cases, the external actuation mechanisms are collarswrapping around at least a portion of the pipette head.

In some embodiments, a system above, alone or in combination, furthercomprises a plurality of switches, an individual switch having an onposition and an off position, wherein the on position permits theplunger associated with the individual switch to move in response tomovement by the motor, and wherein the off position does not permit theplunger associated with the individual switch to move in response tomovement by the motor. In some cases, the switch is a solenoid. In somecases, the switch is operated by a cam operably linked to an additionalmotor.

In some embodiments, a system above, alone or in combination, has atleast one tip mechanism. The at least one tip removal mechanism iswithin a pipette head and is configured to be movable within the pipettehead. In some cases, the at least one tip removal mechanism is externalto the pipette head. In some situations, the at least one tip removalmechanism is a collar wrapping around at least a portion of the pipettehead. In some cases, the pipette head is capable of aspirating and/ordispensing at least 150 uL.

In some embodiments, a system above, alone or in combination, has afluid handling system. The fluid handling apparatus has a height whichdoes not exceed 1 cm, or 2 cm, or 3 cm, or 4 cm, or 5 cm, or 6 cm, or 7cm, or 8 cm, or 9 cm, or 10 cm.

In some embodiments, a system above, alone or in combination, includes aplurality of plungers. At least one plunger is within a pipette head ofsaid plurality, and is configured to be movable within the pipette head.In some cases, the plurality of plungers are independently movable.

In some embodiments, a system above, alone or in combination, has afluid handling apparatus that is capable of dispensing and/or aspiratinga minimum increment of no more than 0.5 uL, or 1 uL.

In some embodiments, a system above, alone or in combination, comprisesa plurality of plungers, wherein at least one plunger is within apipette head of said plurality, and is configured to be movable withinthe pipette head. The plurality of plungers in some cases areindependently movable. In some situations, the system comprises a motorconfigured to permit independent movement of the plurality of plungers.

In some embodiments, an individual pipette head of a plurality ofpipette heads included in a system above is capable of dispensing and/oraspirating 1 uL to 3 mL of fluid.

In some situations, a fluid handling apparatus above, alone or incombination, has a motor (or other actuator) with an axis of rotationthat is horizontal. In some cases, a removable tip of the fluid handlingapparatus is aligned vertically. In some cases, the fluid handlingapparatus comprises at least one plunger within a pipette head of saidplurality, wherein the plunger is configured to be movable within thepipette head; and at least one motor configured to permit movement ofthe plurality of plunger that is not substantially parallel to theremovable tip. In some cases, the plunger is capable of moving in adirection that is substantially perpendicular to the removable tip. Insome situations, the plunger is capable of moving in a horizontaldirection, and wherein the removable tip is aligned vertically.

In some embodiments, a fluid handling apparatus above comprises a firstsection and a second section. The first section is configured to slidewithin the second section. The fluid handling apparatus may furtherinclude a heat spreader surrounding a plunger of the fluid handlingapparatus.

In some embodiments, a fluid handling apparatus includes at least onepipette head, wherein an individual pipette head comprises a pipettenozzle configured to connect with a removable tip, wherein said at leastone pipette head has a fluid path of a given length that terminates atthe pipette nozzle, and wherein the length of the fluid path isadjustable without affecting movement of fluid from the tip when the tipand the pipette nozzle are engaged.

The pipette nozzle may be movable relative to a base operably connectedto the at least one pipette head, thereby adjusting the fluid pathlength. In some cases, the fluid path is formed using rigid components.The fluid path in some cases is formed without the use of flexiblecomponents

In some situations, the fluid handling apparatus further comprises aventilation port within the pipette head. The ventilation port iscapable of having an open position and a closed position. In some cases,a ventilation solenoid determines whether the ventilation port is in theopen position or the closed position. A valve may determine whether theventilation port is in the open position or the closed position. Thevalve can be duty-cycled with periods of less than or equal to 50 ms.

In some situations, the ventilation port is coupled to a positivepressure source that is useful for the expulsion of the fluid. Theventilation port may be coupled to a negative pressure source that isuseful for the aspiration of the fluid.

In some situations, the ventilation port is coupled to atmosphericconditions. The ventilation port may be coupled to a reversible pumpcapable of delivering positive or negative pressure. The pressure sourceis capable of delivering the positive or negative pressure for anextended period of time. In some cases, the removable tip comprises twoopenings, each of which has an embedded passive valve. In somesituations, the embedded passive valves are configured to permit fluidto flow in one direction through a first opening, through a tip body,and through a second opening.

In some situations, at least a 2 cm vertical difference exists betweenthe retracted position and the extended position.

In some embodiments, the pipette nozzle is movable relative to the atleast one plunger. In some situations, adjusting the pipette nozzlebetween the retracted position and the extended position changes a fluidpath length terminating at the pipette nozzle. The fluid path is formedusing only rigid components.

In some embodiments, the plunger comprises a first section and a secondsection wherein at least a portion of the first section is within thesecond section when the pipette nozzle is in the retracted position, andwherein the first section is not within the second section when thepipette nozzle is in the extended position.

In some embodiments, a method above, alone or in combination, comprisesextending a pipette nozzle relative to the base prior to and/orconcurrently with dispensing and/or aspirating a fluid with the tip.

In some embodiments, a method of fluid handling comprises providing atleast one pipette head operably connected to a base, wherein anindividual pipette head comprises a pipette nozzle configured to connectwith a removable tip; providing at least one plunger within a pipettehead of said plurality, wherein the plunger is configured to be movablewithin the pipette head; and retracting the pipette nozzle relative tothe base in first direction prior to and/or concurrently withtranslating the pipette head in a second direction substantiallynon-parallel to the first direction. The first direction and the seconddirection may be substantially perpendicular. In some cases, the firstdirection is a substantially vertical direction while the seconddirection is a substantially horizontal direction.

In some embodiments, a method of fluid handling comprises providing atleast one pipette head operably connected to a base, wherein anindividual pipette head comprises a pipette nozzle configured to connectwith a removable tip; retracting and/or extending the pipette nozzlerelative to the base; and dispensing and/or aspirating a fluid with thetip during said retracting and/or extending. In some situations, themethod further comprises providing at least one plunger within a pipettehead of said plurality, wherein the plunger is configured to be movablewithin the pipette head and/to effect said dispensing and/or aspirating.In some situations, the method further comprises providing a motorcausing the at least one plunger to move within the pipette head. Insome cases, the base supports the at least one pipette head. In somesituations, the pipette nozzle is slidable in a linear direction. Thepipette nozzle may retract and/or extends in a vertical directionrelative to the base.

In some embodiments, a fluid handling apparatus includes a first pipettehead and a second pipette head. In some cases, the first pipette head isa positive displacement pipette head, and the second pipette head is anair displacement pipette head.

In some embodiments, a method for transporting components within adevice comprises providing a plurality of pipette heads, wherein anindividual pipette head comprises a pipette nozzle configured to connectwith a removable tip, wherein the individual pipette head is capable ofdispensing and/or aspirating a fluid with the tip; engaging a sampleprocessing component using at least one pipette head of said plurality;and transporting the sample processing component using at least onepipette head of said plurality. In some cases, the sample processingcomponent is a sample preparation unit or a component thereof, an assayunit or a component thereof, and/or a detection unit or a componentthereof. In some situations, the sample processing component is asupport for a plurality of removable tips and/or vessels. In some cases,the hardware component is picked up using a press-fit between one ormore of the pipette heads and a feature of the hardware component. Insome cases, the hardware component is picked up using a suction providedby one or more of the pipette heads and a feature of the hardwarecomponent.

In some embodiments, a fluid handling apparatus comprises a removabletip; and at least one pipette head, wherein an individual pipette headcomprises a pipette nozzle configured to connect with the removable tip,wherein the apparatus is operably connected to a light source thatprovides light into the tip. In some cases, the tip forms a wave guidecapable of providing a light through the tip to a fluid containedtherein, or capable of transmitting an optical signal from the fluidthrough the tip. In some situations, the removable tip is formed of anoptically transparent material. In some cases, the fluid handlingapparatus further comprises at least one plunger within a pipette headof said plurality, wherein the plunger is configured to be movablewithin the pipette head. In some cases, the pipette nozzle is formedwith a transparent and/or reflective surface. The light source in somecases is within the apparatus. In an example, the light source is withinat least one pipette head. In some situations, the tip comprises a fiberthat conducts said light. In an example, the fiber is formed of anoptically transparent material. In some situations, the fiber extendsalong the length of the removable tip. In some cases, the fiber optic isembedded within the removable tip.

In some embodiments, a fluid handling apparatus comprises a removabletip; and at least one pipette head, wherein an individual pipette headcomprises a pipette nozzle configured to connect with the removable tip,wherein the apparatus is operably connected to an image capture devicethat is configured to capture an image within and/or through the tip.

In some situations, the image capture device is located within theapparatus. In some cases, the image capture device is located within atleast one pipette head.

In some situations, the image capture device is integrally formed withthe apparatus. In some cases, the image capture device is a camera.

In some situations, the image capture device is capable of capturing anelectromagnetic emission and generating an image along one or more of: avisible spectrum, infra-red spectrum, ultra-violet spectrum, gammaspectrum.

In some situations, the fluid handling apparatus further comprises atleast one plunger within a pipette head of said plurality, wherein theplunger is configured to be movable within the pipette head. The imagecapture device may be located at the end of the plunger. The plunger mayinclude (or be formed of) an optically transmissive material. Theplunger may be made of a transparent material.

In some situations, the pipette nozzle is formed with a transparentand/or reflective surface.

In some situations, the fluid handling apparatus further comprises aprocessor on the apparatus.

In some situations, the fluid handling apparatus further comprises aprocessor on the image capture device.

In some embodiments, a fluid handling apparatus comprises a removabletip; at least one pipette head, wherein an individual pipette headcomprises a pipette nozzle configured to connect with the removable tip;and a processor operably connected to the removable tip and/or the atleast one pipette head, wherein the apparatus is configured to varyand/or maintain the position of the removable tip based on instructionsfrom the processor.

In some situations, the removable tip comprises the processor. In somecases, the at least one pipette head comprises the processor. In someimplementations, a first processor of a first removable tip of theapparatus is in communication with a second processor of a secondremovable tip.

In some embodiments, a fluid handling apparatus comprises a movablesupport structure; a plurality of pipette heads sharing the movablesupport structure, wherein an individual pipette head comprises apipette nozzle configured to connect with a removable tip, wherein theplurality of pipette heads are less than or equal to 4 mm apart fromcenter to center.

In some situations, the fluid handling apparatus further comprises aplurality of plungers, wherein at least one plunger is within a pipettehead of said plurality, and is configured to be movable within thepipette head.

In some situations, the fluid handling apparatus further comprises aplurality of transducer driven diaphragms capable of effecting a fluidto be dispensed and/or aspirated through the removable tip.

In some situations, the plurality of pipette heads are movable along thesupport structure so that the lateral distance between the plurality ofpipette heads is variable.

An aspect of the invention provides a method for diagnosing or treatinga subject with the aid of a point of service system, comprising (a)authenticating a subject; (b) obtaining a three-dimensionalrepresentation of the subject with the aid of a three-dimensionalimaging device; (c) displaying the three-dimensional representation to ahealthcare provider in remote communication with the subject, with theaid of a computer system comprising a processor, wherein the system iscommunicatively coupled to the three-dimensional imaging device; and (d)diagnosing or treating the subject with the aid of the displayedthree-dimensional representation of the subject.

Another aspect of the invention provides a point of service system fordiagnosing or treating a subject, comprising a point of service devicehaving a three-dimensional imaging device for providing a dynamicthree-dimensional spatial representation of the subject; and a remotecomputer system being configured to be in communication with thethree-dimensional imaging device and being configured to retrieve thedynamic three-dimensional spatial representation of the subject, whereinthe remote computer system is optionally configured to authenticate thesubject.

An aspect of the invention provides a method for diagnosing or treatinga subject with the aid of a point of care system, comprising:authenticating a subject; obtaining a three-dimensional representationof the subject with the aid of a three-dimensional imaging device;providing the three-dimensional representation to a display of acomputer system of a healthcare provider, the computer systemcommunicatively coupled to the three-dimensional imaging device, thehealthcare provider in remote communication with the subject; anddiagnosing or treating the subject with the aid of the three-dimensionalrepresentation on the display of the computer system.

An additional aspect of the invention provides a point of service systemfor diagnosing or treating a subject, comprising: a point of servicedevice having a three-dimensional imaging device for providing a dynamicthree-dimensional spatial representation of the subject; and a remotecomputer system in communication with the three-dimensional imagingdevice, the remote computer system for authenticating the subject and,subsequent to said authenticating, retrieving the dynamicthree-dimensional spatial representation of the subject.

Additionally, aspects of the invention may be directed to a method formeasuring the body-fat percentage of a subject, comprising: providing apoint of service device having a touchscreen; placing a first finger ona first side of the touchscreen and a second finger on a second side ofthe touchscreen; directing a current from the point of service throughthe body of the subject, wherein the current is directed through thebody of the subject through the first finger and the second finger; anddetermining a body-fat percentage of the subject by measuring theresistance between the first finger and the second finger with the aidof the current directed through the body of the subject.

A method for diagnosing a subject may be provided in accordance withanother aspect of the invention, said method comprising: providing apoint of service device having a touchscreen; placing a first finger ona first side of the touchscreen and a second finger on a second side ofthe touchscreen; directing a current from the point of service throughthe body of the subject, wherein the current is directed through thebody of the subject through the first finger and the second finger;measuring a resistance between the first finger and the second fingerwith the aid of the current directed through the body of the subject;and diagnosing the subject based on the measured resistance.

In some embodiments, a method above, alone or in combination, comprisesputting the subject in contact with a healthcare provider selected bythe subject.

In some cases, diagnosing or treating the subject comprises putting thesubject in contact with the subject's health care provider. In somesituations, diagnosing comprises providing a diagnosis in real-time.

In some embodiments, the three-dimensional imaging device is part of apoint of service system.

In some embodiments, a method above, alone or in combination, furthercomprises identifying the subject prior to diagnosing or treating.

In some embodiments, a method above, alone or in combination, comprisesidentifying a subject by verifying a fingerprint of the subject.

In some embodiments, a method above, alone or in combination, comprisesdiagnosing or treating a subject using a touchscreen display.

In some cases, diagnosing or treating comprises collecting a sample froma subject. The sample in some cases is collected from the subject at thelocation of a healthcare provider. The sample may be collected from thesubject at the location of the subject.

In some situations, a point of service system comprises an imagerecognition module for analyzing at least a portion of the dynamicthree-dimensional spatial representation of the subject for treatment.In some cases, authenticating is performed with the aid of one or moreof a biometric scan, the subject's insurance card, the subject's name,the subject's driver's license, an identification card of the subject,an image of the subject taken with the aid of a camera in the point ofcare system, and a gesture-recognition device.

In some embodiments, a method above, alone or in combination, comprisesdiagnosing a subject by putting the subject in contact with a healthcare provider selected by the subject.

In some embodiments, a method above, alone or in combination, furthercomprises combining a three-dimensional representation of a subject withsubject-specific information. The combination may be made with the aidof a processor. In some cases, the point of service system comprises animage recognition module for analyzing at least a portion of the dynamicthree-dimensional spatial representation of the subject for treatment.

In some cases, a system comprises a touchscreen. The touchscreen may be,for example, a capacitive touchscreen or resistive touchscreen. In somesituations, the touchscreen is at least a 60-point touchscreen.

In some embodiments, for one or more methods above or other methodsprovided herein, the first finger is on a first hand of the subject andthe second finger is on a second hand of the subject.

In some embodiments, a method above, alone or in combination, comprisesdiagnosing a subject by providing a body-fat percentage of the subject.

In accordance with an aspect of the invention, a vessel may comprise: abody configured to accept and confine a sample, wherein the bodycomprises an interior surface, an exterior surface, an open end, and atapered closed end, wherein the vessel is configured to engage with apipette and comprises a flexible material extending across the open endand having a slit/opening that is configured to prevent fluid frompassing through the flexible material in the absence of an objectinserted through the slit/opening.

Aspects of the invention may be directed to a vessel, comprising: a bodyconfigured to accept and confine a sample of no more than about 100 μL,wherein the body comprises an interior surface, an exterior surface, andan open end, wherein the vessel comprises a flexible material extendingacross the open end and having a slit/opening that is configured toprevent fluid from passing through the flexible material in the absenceof an object inserted through the slit/opening.

A vessel may be provided in accordance with additional aspect of theinvention, said vessel comprising: a body configured to accept andconfine a sample, wherein the body comprises an interior surface, anexterior surface, a first end, a second end, and a passage between thefirst end and the second end, wherein the vessel comprises a materialextending across the passage capable of having (1) molten state that isconfigured to prevent fluid from passing through the material in theabsence of an object inserted through the material, and (2) a solidstate that is configured to prevent fluid and the object from passingthrough the material.

Also, aspects of the invention may provide an injection molding templatecomprising a substrate comprising a planar surface and a plurality ofprojections; and an opposing mold comprising a plurality of indentationswherein the projections are configured to be positionable within theindentations, wherein an individual projection of said pluralitycomprises a cylindrical portion of a first diameter, and a funnel shapedportion contacting the cylindrical portion, wherein one end of thefunnel shaped portion contacting the cylindrical portion has the firstdiameter, and a second end of the funnel shaped portion contacting theplanar surface has a second diameter.

In accordance with an additional aspect of the invention, a system maycomprise: a vessel configured to accept and confine a sample, whereinthe vessel comprises an interior surface, an exterior surface, an openend, and an opposing closed end; and a tip configured to extend into thevessel through the open end, wherein the tip comprises a first open endand second open end, wherein the second open end is inserted into thevessel, wherein the vessel or the tip further comprises a protrudingsurface feature, optionally at or near the closed end, that prevents thesecond open end of the tip from contacting the bottom of the interiorsurface of the closed end of the vessel.

In some embodiments, a vessel provided above or elsewhere hereinincludes flexible material. In some cases, the flexible material is amembrane. In some cases, the flexible material is formed from asilicon-based material.

In some embodiments, a vessel provided above or elsewhere hereinincludes a cap configured to contact the body at the open end, whereinat least a portion of the cap extends into the interior of the body. Insome cases, the cap comprises a passageway through which the flexiblematerial extends.

In some embodiments, a vessel provided above or elsewhere hereinincludes a body that has a cylindrical portion of a first diameterhaving an open end and a closed end, and a funnel shaped portionedcontacting the open end, wherein one end of the funnel shaped portioncontacting the open end has a first diameter, and a second end of thefunnel shaped portion has a second diameter. In some cases, the seconddiameter is less than the first diameter. In other cases, the seconddiameter is greater than the first diameter. In other cases, the seconddiameter is equal to the first diameter. In some cases, the second endof the funnel shaped portion is configured to engage with a removablecap.

In some embodiments, a vessel provided above or elsewhere hereinincludes a flexible material that is a membrane. The flexible material,in some cases, is formed from a silicon-based material.

In some embodiments, a vessel provided above or elsewhere hereinincludes a cap configured to contact the body at the open end, whereinat least a portion of the cap extends into the interior of the body. Insome cases, the cap comprises a passageway through which the flexiblematerial extends.

In some embodiments, a vessel provided or elsewhere herein has a bodythat has a cylindrical portion of a first diameter having an open endand a closed end, and a funnel shaped portioned contacting the open end,wherein one end of the funnel shaped portion contacting the open end hasa first diameter, and a second end of the funnel shaped portion has asecond diameter. In some cases, the second diameter is less than thefirst diameter. In other cases, the second diameter is greater than thefirst diameter. In some situations, the second end of the funnel shapedportion is configured to engage with a removable cap.

In some embodiments, a vessel provided above or elsewhere hereincomprises a material extending across the passage capable of having (1)molten state that is configured to prevent fluid from passing throughthe material in the absence of an object inserted through the material,and (2) a solid state that is configured to prevent fluid and the objectfrom passing through the material. In some cases, the material is a wax.In some cases, the material has a melting point between about 50° C. and60° C. In some situations, the object is capable of being insertedthrough the material and removed from the material while the material isin the molten state. In some cases, the material is configured to allowsaid object to be inserted into the material and removed from thematerial while the material is in the molten state. In some embodiments,at least a portion of the object is coated with the material when theobject is removed from the material.

In some embodiments, an injection molding template comprises a substratecomprising a planar surface and a plurality of projections; and anopposing mold comprising a plurality of indentations wherein theprojections are configured to be positionable within the indentations,wherein an individual projection of said plurality comprises acylindrical portion of a first diameter, and a funnel shaped portioncontacting the cylindrical portion, wherein one end of the funnel shapedportion contacting the cylindrical portion has the first diameter, and asecond end of the funnel shaped portion contacting the planar surfacehas a second diameter. The plurality of projections in some cases arearranged in an array. In some situations, the volume of the projectionsis less than or equal to 100 microliters (‘uL”), 50 uL, 20 uL, 10 uL, or1 uL. In some cases, the indentations comprise a cylindrical portion anda funnel shaped portioned contacting the cylindrical portion.

In some embodiments, a system provided above, alone or in combination,such as a vessel, includes surface features that are integrally formedon the bottom interior surface of the vessel. In some embodiments, thesurface features are a plurality of bumps on the bottom interior surfaceof the vessel.

In some embodiments, an apparatus provided above, alone or incombination, comprises a planar substrate comprising a plurality ofdepressions; and a plurality of tips of having a configuration providedabove or elsewhere herein, wherein the tips are at least partiallyinserted into the plurality of depressions and supported by thesubstrate. In some cases, the apparatus forms a microtiter plate.

In some aspects of the invention, a centrifuge may be provided, saidcentrifuge comprising: a base having a bottom surface, said base beingconfigured to rotate about an axis orthogonal to the bottom surface,wherein the base comprises one or more wing configured to fold over anaxis extending through the base, wherein a wing comprises an entireportion of base on a side of the axis, wherein the wing comprises acavity to receive a sample vessel, wherein the sample vessel is orientedin a first orientation when the base is at rest, and is configured to beoriented at a second orientation when the base is rotating.

A centrifuge comprise, in accordance with an aspect of the invention, abase having a bottom surface and a top surface, said base beingconfigured to rotate about an axis orthogonal to the bottom surface,wherein the base comprises one or more bucket configured to pivot abouta pivot axis, configured to permit at least a portion of the bucket topivot upwards past the top surface, and wherein the bucket comprises acavity to receive a sample vessel, wherein the cavity is configured tobe oriented in a first orientation when the base is at rest, and isconfigured to be oriented at a second orientation when the base isrotating.

Additionally, aspects of the invention may be directed to a centrifugecomprising: a base having a bottom surface and a top surface, said basebeing configured to rotate about an axis orthogonal to the bottomsurface, wherein the base comprises one or more bucket configured topivot about a pivot axis, and said bucket is attached to a weightconfigured to move in a linear direction, thereby causing the bucket topivot, and wherein the bucket comprises a cavity to receive a samplevessel, wherein the cavity is configured to be oriented in a firstorientation when the base is at rest, and is configured to be orientedat a second orientation when the base is rotating.

In accordance with another aspect of the invention, a centrifuge maycomprise: a brushless motor assembly comprising a rotor configured torotate about a stator about an axis of rotation; and a base comprisingone or more cavities configured to receive one or more fluidic samples,said base affixed to the rotor, wherein the base rotates about thestator and a plane orthogonal to the axis of rotation of the brushlessmotor is coplanar with a plane orthogonal to the axis of rotation of thebase.

Aspects of the invention may be directed to, a centrifuge comprising: abrushless motor assembly comprising a rotor configured to rotate about astator about an axis of rotation, wherein the brushless motor has aheight in the direction of the axis of rotation; and a base comprisingone or more cavities configured to receive one or more fluidic samples,said base affixed to the rotor, wherein the base rotates about thestator and said base has a height in the direction of the axis ofrotation, and wherein the height of the brushless motor assembly is nogreater than twice the height of the base.

A system may be provided in accordance with another aspect of theinvention, said system comprising: at least one module mounted on asupport structure, wherein said at least one module comprises a samplepreparation station, assay station, and/or detection station; and acontroller operatively coupled to said at least one module and anelectronic display, said electronic display having a graphical userinterface (GUI) for enabling a subject to interact with the system,wherein the system is configured to perform (a) at least one samplepreparation procedure selected from the group consisting of sampleprocessing, centrifugation, magnetic separation, and chemicalprocessing, and (b) multiple types of assays selected from the groupconsisting of immunoassay, nucleic acid assay, receptor-based assay,cytometric assay, colorimetric assay, enzymatic assay, electrophoreticassay, electrochemical assay, spectroscopic assay, chromatographicassay, microscopic assay, topographic assay, calorimetric assay,turbidimetric assay, agglutination assay, radioisotope assay,viscometric assay, coagulation assay, clotting time assay, proteinsynthesis assay, histological assay, culture assay, osmolarity assay,and combinations thereof.

In some embodiments, assays described above or elsewhere herein may bemeasured at the end of the assay (an “end-point” assay) or at two ormore times during the course of the assay (a “time-course” or “kinetic”assay).

Aspects of the invention may be directed to a system, comprising: asupport structure having a mounting station configured to support amodule among a plurality of modules, an individual module configured toperform (i) at least one sample preparation procedure selected from thegroup consisting of sample processing, centrifugation, magneticseparation, and/or (ii) at least one type of assay selected from thegroup consisting of immunoassay, nucleic acid assay, receptor-basedassay, cytometric assay, colorimetric assay, enzymatic assay,electrophoretic assay, electrochemical assay, spectroscopic assay,chromatographic assay, microscopic assay, topographic assay,calorimetric assay, turbidimetric assay, agglutination assay,radioisotope assay, viscometric assay, coagulation assay, clotting timeassay, protein synthesis assay, histological assay, culture assay,osmolarity assay, and combinations thereof; a controller operativelycoupled to said plurality of modules, wherein the controller isconfigured to provide one or more instructions to said module orindividual modules of said plurality of modules to facilitateperformance of the at least one sample preparation procedure or the atleast one type of assay; and an electronic display operatively coupledto said controller, said electronic display having a graphical userinterface (GUI) for enabling a subject to interact with the system.

Systems above or elsewhere herein, alone or in combination, may comprisea plurality of modules mounted on the support structure, an individualmodule of said plurality of modules comprising a sample preparationstation, assay station and/or detection station. An individual modulemay be configured to perform said at least one sample preparationprocedure and/or said at least one type of assay without the aid ofanother module in said systems above or elsewhere herein, alone or incombination.

In some systems above or elsewhere herein, alone or in combination, acontroller may be mounted on the support structure.

The GUI provided in systems above or elsewhere herein, alone or incombination, may be configured to provide a guided questionnaire to saidsubject.

The guided questionnaire may comprise one or more graphical and/ortextual items, in systems above or elsewhere herein, alone or incombination. In some embodiments, the guided questionnaire may beconfigured to collect, from said subject, information selected from thegroup consisting of dietary consumption, exercise, health condition andmental condition.

In the systems above or elsewhere herein, alone or in combination, anelectronic display may be mounted on the support structure. In someembodiments, the electronic display may be mounted on a supportstructure of a remote system, such as systems above or elsewhere herein,alone or in combination. In accordance with some embodiments of theinvention, the electronic display may be an interactive display. Insystems above or elsewhere herein, alone or in combination, aninteractive display may be a capacitive-touch or resistive-touchdisplay.

A communications module may be operatively coupled to said controller,the communications module for enabling the system to communicate with aremote system, which may include systems above or elsewhere herein,alone or in combination.

Systems above or elsewhere herein, alone or in combination, may furthercomprise a database operatively coupled to the controller, said databasefor storing information related to said subject's dietary consumption,exercise, health condition and/or metal condition.

Optionally, one may use paper-based system that becomes colored (blotreaction down) and does a colorimetric assay on the paper, measuringreflectance, instead of a system that uses transmission through asample.

Other detection methods may include detecting agglutination where thesystem uses imaging from imaging device(s) in the system. Turbidimetricmeasurement techniques can use the spectrophotometer as the detector.

Optionally, the system may run or measure a coagulation assay on anucleic acid assay station and there may be non-cytometry assay run ormeasured in the cytometer module.

Optionally, the system may measure lead or other metals that complexwith porphyrins and result in a wavelength shift. In the event of awavelength shift when a metal complexes with porphyrin, this may bedetectable by spectrophotometery or other techniques for detecting thewavelength shift.

Optionally, the system may have a detector that measures heat in thesample.

Optionally, chromatographic techniques may be used to detect generalchemistry assays. HPLC may be used. The sample may be processed so thatits analyte levels are measured by UV or fluorescence. Some embodimentsmay use a filter that facilitates chromatography as the system doesseparations on the sample, such as in a tip.

Optionally, general chemistry assays may be characterized as an assay ona non-phase separated sample, wherein there is no washing or removalstep to removal sample. The assays may occur in the homogenous phaseversus the heterogeneous phase. The samples may be processed inadditive, non-separating type of manner Separating steps for assays notin the general chemistry group of assays may involve washing of beads,removing reaction medium to add new medium. In one non-limiting example,the assays in the general chemistry group are primarily not binder orantibody based. Typically, the assays in this group do not involveamplification of nucleic acids, imaging cells on a microscopy stage, orthe determination of analyte level(s) in solution based on a labeledantibody or binder.

In some embodiments, provided herein is a biological sample processingdevice comprising: a) a sample handling system; b) a detection station;c) a cytometry station comprising an imaging device and a stage forreceiving a microscopy cuvette; and d) an assay station configured tosupport multiple components comprising i) a biological sample and ii) atleast a first, a second, and a third fluidically isolated assay unit,wherein the sample handling system is configured to i) transfer at leasta portion of the biological sample to the first assay unit, the secondassay unit, and the third assay unit; ii) transfer the first and secondassay units containing biological sample to the detection station; andiii) transfer the third assay unit containing biological sample to thecytometry station.

In some embodiments, provided herein is a biological sample processingdevice comprising: a) a sample handling system; b) a detection station;c) a cytometry station comprising an imaging device and a stage forreceiving a microscopy cuvette; and d) an assay station configured tosupport multiple components comprising i) a biological sample, ii) atleast a first, a second, and a third fluidically isolated assay unit,and iii) reagents to perform A) at least one immunoassay; B) at leastone general chemistry assay; and C) at least one cytometry assay, andwherein the sample handling system is configured to i) transfer at leasta portion of the biological sample to the first assay unit, the secondassay unit, and the third assay unit; ii) transfer the first and secondassay units containing biological sample to the detection station; andiii) transfer the third assay unit containing biological sample to thecytometry station.

In some embodiments, provided herein is a biological sample processingdevice comprising: a) a sample handling system; b) a first detectionstation comprising an optical sensor; c) a second detection stationcomprising a light source and an optical sensor; d) a cytometry stationcomprising an imaging device and a stage for receiving a microscopycuvette; and e) an assay station configured to support i) a biologicalsample, ii) at least a first, a second, and a third fluidically isolatedassay unit, and iii) reagents to perform A) at least one luminescenceassay; B) at least one absorbance, turbimetric, or colorimetric assay;and C) at least one cytometry assay; wherein the first assay unit isconfigured to perform a luminescence assay, the second assay unit isconfigured to perform an absorbance, turbidimetric, or colorimetricassay, the third assay unit is configured to perform a cytometry assay,and the sample handling system is configured to i) transfer at least aportion of the biological sample to the first, second, and third assayunits; ii) transfer the first assay unit containing biological sample tothe first detection station; iii) transfer the second assay unitcontaining biological sample to the second detection station; and iv)transfer the third assay unit containing biological sample to the stageof the cytometry station.

In some embodiments, provided herein is biological sample processingdevice, comprising: a) a sample handling system; b) a detection stationcomprising an optical sensor; c) a fluidically isolated samplecollection unit configured to retain a biological sample; d) an assaystation comprising at least a first, second, and third fluidicallyisolated assay unit, wherein the first unit comprises an antibody, thesecond unit comprises an oligonucleotide, and the third unit comprises achromogenic substrate; and e) a controller, wherein the controller isoperatively coupled to the sample handling system, wherein the samplehandling system is configured to transfer a portion of the biologicalsample from the sample collection unit to each of the first assay unit,the second assay unit, and the third assay unit, and the device isconfigured to perform an immunoassay, a nucleic acid assay, and ageneral chemistry assay comprising a chromogenic substrate.

In some embodiments, provide herein is a biological sample processingdevice, comprising a housing containing therein: a) a sample handlingsystem; b) a detection station comprising an optical sensor; c) afluidically isolated sample collection unit configured to retain abiological sample; d) an assay station comprising at least a first,second, and third fluidically isolated assay unit, wherein the firstunit comprises a first reagent, the second unit comprises a secondreagent, and the third unit comprises a third reagent; and e) acontroller, wherein the controller comprises a local memory and isoperatively coupled to the sample handling system and the detectionstation; wherein the device is configured to perform assays with any oneor more of the first, second, and third assay units; wherein the localmemory of the controller comprises a protocol comprising instructionsfor: i) directing the sample handling system to transfer a portion ofthe biological sample to the first assay unit, the second assay unit andthe third assay unit; and ii) directing the sample handling system totransfer the first unit, the second unit, and the third assay unit tothe detection station.

In some embodiments, provided herein is a method of performing at least4 different assays selected from immunoassays, cytometric assays, andgeneral chemistry assays on a biological sample, the method comprising:a) introducing a biological sample having a volume of no greater than 2ml, 1 ml, 500 microliters, 300 microliters, 200 microliters, 100microliters, 50 microliters, 25 microliters, 25 microliters, 10microliters, or 5 microliters into a sample processing device, whereinthe device comprises: i) a sample handling system; ii) a detectionstation; iii) a cytometry station comprising an imaging device and astage for receiving a microscopy cuvette; and iv) an assay stationcomprising at least a first, a second, a third, a fourth, and a fifthindependently movable assay unit; b) with the aid of the sample handlingsystem, transferring a portion of the biological sample to each of thefirst, second, third, and fourth assay units, wherein a different assayis performed in each of the first, second, third, and fourth assayunits; c) with the aid of the sample handling system, transferring thefirst, second, third, and fourth assay units to the detection station orcytometry station, wherein assay units comprising immunoassays orgeneral chemistry assays are transferred to the detection station andassay units comprising cytometric assays are transferred to thecytometry station; d) with the aid of the detection station or cytometrystation, obtaining data measurements of the assay performed in each ofthe first, second, third, and fourth assay units. In some embodiments,the above method may apply to a method of performing 2, 3, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100,150, 200, or more different assays.

In some embodiments, provided herein is a method of processing abiological sample, comprising: a) introducing a sample having a volumeof 2 ml, 1 ml, 500 microliters, 300 microliters, 200 microliters, 100microliters, 50 microliters, 25 microliters, 25 microliters, 10microliters, or 5 microliters or less into a sample processing devicecomprising i) a sample handling system; ii) at least a first and asecond fluidically isolated vessel; and iii) a diluent, wherein thesample comprises bodily fluid at a first concentration; b) with the aidof the sample handling system, mixing at least a portion of the samplewith the diluent to generate a diluted sample, wherein the dilutedsample comprises bodily fluid at a second concentration, and the secondconcentration of bodily fluid is one-half, one-third, one-quarter,one-tenth, or less of the first concentration of bodily fluid; and, c)with the aid of the sample handling system, transferring at least aportion of the diluted sample to the first and the second fluidicallyisolated vessels.

In some embodiments, provided herein is a method of processing abiological sample, comprising: a) introducing a sample having a volumeof 2 ml, 1 ml, 500 microliters, 300 microliters, 200 microliters, 100microliters, 50 microliters, 25 microliters, 25 microliters, 10microliters, or 5 microliters or less into a sample processing devicecomprising i) a sample handling system; ii) at least a first and asecond fluidically isolated vessel; iii) a diluent; and iv) acentrifuge, wherein the sample comprises bodily fluid at a firstconcentration; b) with the aid of the sample handling system,introducing at least a portion of the sample into the centrifuge; c)centrifuging the sample, to generate a centrifuged sample; d) with theaid of the sample handling system, removing at least a portion of thecentrifuged sample from the centrifuge; and e) with the aid of thesample handling system, mixing at least a portion of the centrifugedsample with the diluent to generate a diluted sample, wherein thediluted sample comprises bodily fluid at a second concentration, and thesecond concentration of bodily fluid is one-half, one-third,one-quarter, one-tenth, or less of the first concentration of bodilyfluid; and, f) with the aid of the sample handling system, transferringat least a portion of the diluted sample to the first and the secondfluidically isolated vessels.

In some embodiments, provided herein is a method of preparing abiological sample, comprising: a) introducing a biological sample and atleast one isolated vessel into a sample processing device comprising acentrifuge and a sample handling system; b) with the aid of the samplehandling system, introducing at least a portion of the biological sampleinto the centrifuge, wherein the centrifuge comprises one or morecavities and wherein the one or more cavities are configured to receivea total of no more than 2 ml, 1.5 ml, 1 ml, 750 microliters, 500microliters, 300 microliters, 200 microliters, 100 microliters, 50microliters, 25 microliters, 25 microliters, or 10 microliters betweenall of the one or more cavities; c) centrifuging the sample, to generatea centrifuged sample; d) with the aid of the sample handling system,removing at least a portion of the centrifuged sample from thecentrifuge; and e) with the aid of the sample handling system,transferring centrifuged sample removed from the centrifuge from step d)into the fluidically isolated vessel.

In some embodiments, in an assay station described above or elsewhereherein, the assay station is configured to support multiple components.The components may comprise, for example, any one or more of: i) abiological sample; ii) any number of fluidically isolated assay units(for example, at least a first, a second, and a third fluidicallyisolated assay unit); iii) any number of fluidically isolated reagentunits (its (for example, at least a first, a second, and a thirdfluidically isolated reagent unit); iv) reagents to support any numberof immunoassays; v) reagents to support any number of general chemistryassays; vi) reagents to support any number of cytometry assays; vii)reagents to support any number of nucleic acid assays; viii) reagents toperform any number of luminescent assays; ix) any number of absorbance,turbimetric, or colorimetric assays; x) any number of fluidicallyisolated vessels; or xi) two or more fluidically isolated vessels whichare physically linked.

In some embodiments, an assay station described above or elsewhereherein that is configured to support multiple components may contain thecomponents.

In some embodiments, a sample processing device described above orelsewhere herein that contains an assay station receiving location mayalso contain an assay station. In some embodiments, in a sampleprocessing device described above or elsewhere herein that contains anassay station, the assay station may be located in an assay stationreceiving location. In some embodiments, a sample processing devicedescribed above or elsewhere herein that is configured to receive anassay station may contain an assay station.

In some embodiments, an assay station or cartridge described above orelsewhere herein may be configured to support a biological sample of nogreater than 100 ml, 50 ml, 30 ml, 20 ml, 10 ml, 5 ml, 2 ml, 1.5 ml, 1ml, 750 microliters, 500 microliters, 400 microliters, 300 microliters,200 microliters, 100 microliters 75 microliters, 50 microliters, 40microliters, 30 microliters, 20 microliters, 10 microliters, 5microliters, 3 microliters, or 1 microliter.

In some embodiments, an assay station or cartridge described above orelsewhere herein may be configured to support a sample collection unit.

In some embodiments, a sample handling system described above orelsewhere herein may be configured to any one or more of: i) transfer atleast a portion of a biological sample to or between one or more assayunits, cuvettes, tips, or other vessels; ii) transfer any one or moreassay units, cuvettes, tips, or other vessels between (to or from) anassay station and a detection station; iii) transfer any one or moreassay units, cuvettes, tips, or other vessels between (to or from) anassay station and a cytometry station; iv) transfer any one or moreassay units, cuvettes, tips, or other vessels between (to or from) anassay station and any one or more different detection stations.

In some embodiments, an assay unit described above or elsewhere hereinmay be a cuvette.

In some embodiments, an assay unit described above or elsewhere hereinmay be a cytometry cuvette configured to interface with a microscopystage.

In some embodiments, assay units described above or elsewhere herein maybe fluidically isolated.

In some embodiments, assay units described above or elsewhere herein maybe fluidically isolated and independently movable.

In some embodiments, assay units described above or elsewhere herein mayhave at least two different configurations or shapes.

In some embodiments, a sample processing device described above orelsewhere herein may contain a housing. In some embodiments, some or allof the components of the device may be within the device housing.

In some embodiments, a sample processing device or a module describedabove or elsewhere herein may contain, one, two, three, four or moredifferent detection stations. The detection stations may containdifferent types of detection units.

In some embodiments, in a sample processing device or module describedabove elsewhere containing a controller, the controller may beoperatively coupled to any component within the device or module.

In some embodiments, in a sample processing device or module describedabove elsewhere containing a controller, the controller may contain alocal memory.

In some embodiments, in a sample processing device or module describedabove elsewhere containing a controller, the controller may contain aprotocol comprising instructions for directing a sample handling systemto transfer a portion of a biological sample to or from one or morefluidically isolated assay units, tips, cuvettes, or other vessels.

In some embodiments, in a sample processing device or module describedabove elsewhere containing a controller, the controller may beconfigured to direct a sample handling system to transfer a portion of abiological sample to or from one or more fluidically isolated assayunits, tips, cuvettes, or other vessels.

In some embodiments, in a sample processing device or module describedabove elsewhere containing a controller, the controller may contain aprotocol comprising instructions for directing a sample handling systemto transfer one or more fluidically isolated assay units, tips,cuvettes, or other vessels to or from a detection station.

In some embodiments, in a sample processing device or module describedabove elsewhere herein containing a controller, the controller maycontain a protocol comprising instructions for directing a samplehandling system to transfer one or more fluidically isolated assayunits, tips, cuvettes, or other vessels to or from a cytometry station.

In some embodiments, an assay unit described above elsewhere herein maybe configured for interfacing with a spectrophotometer. In someembodiments, assay reagents may be added or mixed in an assay unit orother vessel while the assay unit or other vessel is located in aspectrophotometer.

In some embodiments, a single cartridge described above or elsewhereherein may contain two or more different types of biological sample(e.g. blood, urine, saliva, nasal wash, etc.). In some embodiments, asample processing device described above or elsewhere herein may beconfigured for simultaneously performing assays with two or moredifferent types of biological sample. In some embodiments, a singlecartridge described above or elsewhere herein may contain biologicalsamples from two or more different subjects. In some embodiments, asample processing device described above or elsewhere herein may beconfigured for simultaneously performing assays with biological samplesfrom two or more different subjects.

In one embodiment, the controller may be configured to allow forvariable location tip pickup and/or dropoff. In some embodiments, thecontroller is a programmable circuit that is used to direct a samplehandling system to pickup and dropoff sample devices and/or vessels atfixed locations, such as certain stations that have fixed locations fortheir vessel receiving locations. Some may have a controller that isconfigured to also direct the sample handling system to pickup and/ordropoff devices, vessels, or elements at variable locations, such as butnot limited to a centrifuge vessel where the stopping location of thecentrifuge rotor bucket is variable. In such a non-limiting example, thecentrifuge may have position sensor(s) such as but not limited tooptical and/electrical sensor that can relay to the processor thestopping location of the centrifuge rotor.

In another embodiment, a multi-analysis system is described herein thatcomprises a system that can process at least a certain number ofdifferent types of assays from a single fluid sample. In one embodiment,this fluid sample is about 140 microliters to about 150 microliters ofsample fluid. Optionally, this fluid sample is about 130 microliters toabout 140 microliters. Optionally, this fluid sample is about 120microliters to about 130 microliters. Optionally, this fluid sample isabout 110 microliters to about 130 microliters. Optionally, this fluidsample is about 100 microliters to about 120 microliters. Optionally,this fluid sample is about 90 microliters to about 110 microliters.Optionally, this fluid sample is about 80 microliters to about 100microliters. Optionally, this fluid sample is about 70 microliters toabout 90 microliters. Optionally, this fluid sample is about 60microliters to about 80 microliters. Optionally, this fluid sample isabout 50 microliters to about 70 microliters. Optionally, this fluidsample is about 40 microliters to about 60 microliters. Optionally, thisfluid sample is about 30 microliters to about 50 microliters.Optionally, this fluid sample is about 20 microliters to about 40microliters. Optionally, this fluid sample is about 10 microliters toabout 30 microliters.

In another embodiment, a method is provided of concurrent analysisdifferent assay types in multiple tips, cuvettes, or other samplevessels. As discussed herein, the system can multiplex the analysis ofthe same sample, wherein the same sample is aliquoted into multiplesample aliquots, typically multiple diluted samples. In one non-limitingexample, each of these diluted samples is processed in different samplevessels. The aliquoting may occur without having to pass the samplethrough any tubing wherein sample enters from one end and exits from adifferent end of the tube. This type of “tube” based transport is rifewith dead space sample is often lost during transport, resulting inwasted sample and inaccurate sample volume control.

In yet another embodiment, one example of the system configurationallows for processing, simultaneously or sequentially, of differentsignal types, such as those from an optical domain and those from anon-optical domain such as but not limited to electrochemical or thelike. Optionally, different signal types may also be different types ofoptical signals, but all occurring simultaneously for dilute aliquots ofthe same samples, optionally each a different dilution, optionally eachfor a different assay type, and/or optionally from different shapedsample vessels.

In yet another embodiment, a cartridge is provided that comprisestherein at least three different types of reagents or tips in thecartridge. Optionally, the cartridge comprises at least two differenttypes of reagents and at least two different types of pipette tips.Optionally, the cartridge comprises at least three different types ofreagents and at least two different types of pipette tips or samplevessels. Optionally, the cartridge comprises at least three differenttypes of reagents and at least three different types of pipette tips orsample vessels. Optionally, the cartridge comprises at least fourdifferent types of reagents and at least three different types ofpipette tips or sample vessels. Optionally, the cartridge comprises atleast four different types of reagents and at least four different typesof pipette tips or sample vessels. It should be understood that someembodiments may have the cartridge as a disposable. One embodiment ofthe cartridge may only have different types of pipette tips or samplevessels, but no reagents in the cartridge. One embodiment of thecartridge may only have different types of pipette tips or samplevessels, but no reagents in the cartridge and only diluents. Optionally,some may split the reagents into one cartridge and vessels/tips inanother cartridge (or some combination therein). Optionally, oneembodiment of the cartridge may have different types of pipette tips orsample vessels and a majority but not all of the reagents thereon. Insuch a configuration, the remainder of the reagents may be on thehardware of the device and/or provided by at least another cartridge.Some embodiments may comprise loaded more than one cartridge onto thecartridge receiving location, such as a tray. Optionally, someembodiments may combine two cartridges together and load that joinedcartridge (that may be physically linked) onto the cartridge receivinglocation. Optionally, in one embodiment, a majority of reagents forassays are in the device, not the disposable such as the cartridge.Optionally, a majority of physical items such as but not limited totips, vessels, or the like returned to cartridge for disposal.Optionally, prior to ejecting the disposable, the system may move unusedor other fluids in the vessel to absorbent pads or use reagentneutralization prior to disposal, thus minimizing contamination risk.The may involve further diluting any sample, reagent, or the like. Thismay involve using neutralizers or the like to quench or renders harmlessreagents in the cartridge.

In a still further embodiment, the system may comprise a control thatuses a protocol that sets forth processing steps for all of theindividual stations and hardware such as the sample handling system inthe multi-analysis device. By way of non-limiting example, theseprotocols are downloaded from a remote server based on criteria such asbut not limited to cartridge ID, patient ID, or the like. Additionally,prior to cartridge insertion, upon verification of patient ID and/ortest order, the remote server may also perform a translation stepwherein the server or local device can inform the local operator whichcartridge to selected based on the requested combination of testsassociated with the patient ID, lab order, or other information. Thiscan be of particular use as this translation step can in one embodimentaccount for cartridges in inventory at the remote location and thatbecause each cartridge is a multi-assay type cartridge, it is notobvious which cartridge should be selected, unlike known cartridges thatonly perform one assay per cartridge. Here, because of the multi-assayper cartridge, some embodiments may have multiple cartridges that canperform some or all of the requested test and a weighing of inventory,maximizing utilization of cartridge reagents, and/or minimizingcartridge cost can be factored into the cartridge that the system asksthe local user to insert into the system.

In a still further embodiment, the system is deployable in manylocations in the sense that the local operator has limited in whatoperations the local user can control on the device. By way ofnon-limiting example, the user can only select which cartridge isinserted into the sample processing device. In this example, the userdoes not directly do any sample pipetting or the like. The user caninsert sample vessels onto the cartridge and then insert the cartridgeinto the device. Error checking algorithm can determine if the userinserted the correct cartridge for the subject sample based on IDinformation on the cartridge and/or the sample vessel.

In a still further embodiment, a system and method is provided forperforming multiple assays from a single sample, where original sampleis less than a certain volume of sample (in one nonlimiting example, nomore than about 200 microliters). In this example, the dilution of thesample to aliquot the same sample is variable, not fixed, and is basedon the assays to be run. In one embodiment, a system and method isprovided so that whole blood, serum, and plasma can be extracted fromthe single sample of less than 200 microliters. In one embodiment, asystem and method is provided so that whole blood, serum, plasma, andcells can be extracted from the single sample of less than 200microliters. “Sample” division/cell separation steps is one factor thatcan enable multi-testing using such reduced starting sample volume. Inone embodiment, at least 40 assays are run on sample extracted from nomore than about 200 microliters of original undiluted sample.Optionally, at least 20 assays are run on sample extracted from no morethan about 150 microliters of original undiluted sample. Optionally, atleast 20 assays are run on sample extracted no more than about 100microliters of original undiluted sample. Optionally, at least 20 assaysare run on sample extracted from no more than about 80 microliters oforiginal undiluted sample. Optionally, at least 20 assays are run onsample extracted from no more than about 60 microliters of originalundiluted sample. Optionally, at least 20 assays are run on sampleextracted from no more than about 40 microliters of original undilutedsample. Optionally, at least 20 assays are run on sample extracted fromno more than about 30 microliters of original undiluted sample.

In a still further embodiment, a system and method is provided whereintest results are completed within one hour, prior to start of testing,there is real-time insurance verification to determine cost to thesubject to run the test. Herein, testing comprises at least 10 assaysare run on sample extracted from no more than about 150 microliters oforiginal undiluted sample. Optionally, testing comprises at least 10assays are run on sample extracted from no more than about 100microliters of original undiluted sample

In a still further embodiment, a system and method wherein the samehardware system can measure analytes or other characteristics of urine,blood, and feces all by using same hardware, but different disposablesuch as a cartridge.

In one embodiment, a method is provided for evaluating a biologicalsample collected from a subject, said method comprising: providing acartridge having a plurality of receptacles; loading the sample into oneor more receptacles; delivering a loaded cartridge to an analyzer havinga processor, one or more sensors, and a fluid handling system, thecartridge having a multiplicity of selected reagents into receptacles,said reagents being sufficient to perform at least two assays selectedfrom a group of at least 10 assays; wherein the processor is configuredto control the fluid handling system and the sensors to react the samplewith the reagents to perform the at least two assays. Optionally, in thecartridge, substantially of the all of the reagents to be used in theassays for that cartridge are in the cartridge. By substantially, oneembodiment means the volume of reagent. Optionally, by substantially,another embodiment means the types of reagents.

It should be understood that any of the foregoing may be configured tohave one or more of the following features. By way of non-limitingexample, one embodiment may have a method wherein loading the samplecomprises loading sample fluid from one subject and one subject only.Optionally, the method further comprising pretreating the sample toproduce at least two samples which have been pretreated differently andwhich are loaded into separate receptacles. Optionally, at least twosamples are pretreated with different anti-coagulants. Optionally, thesample is held in a holder and the holder is loaded into a receptacle.Optionally, the fluid handling system comprises at least one pipettewhich transfers the sample and the reagents among reaction zones andtest zones. Optionally, the reagents are selected to perform bothprimary assays and reflex assays. Optionally, the processor isconfigured to perform one or more reflex assays if the results of aprimary assay are out of a normal range. Optionally, the cartridge isencoded with information which defines the assays to be performed.Optionally, the cartridge is encoded with information which defines thelocations of the reagents.

In another embodiment, a method is provided for evaluating a biologicalsample from a single subject, the method comprising: dividing the sampleinto multiple aliquots; pretreating such aliquot wherein at least twoaliquots are pretreated differently; delivering each of the pretreatedaliquots to an analyzer having reagents selected to perform at least twoassays selected from a group of at least 10 assays, a processor, aplurality of sensors sufficient to perform each of said at least 10assays, and a fluid handling system; wherein the processor is adapted tocontrol the fluid handling system and the sensors to react each of thesamples with reagents and analyze the reacted samples with sensor(s)selected to perform the at least two assays.

It should be understood that any of the foregoing may be configured tohave one or more of the following features. By way of non-limitingexample, one method of pretreating comprises treating at least onealiquot with a first anti-coagulant and at least one aliquot with asecond anti-coagulant. Optionally, the sensors include at least twosensors selected from the group consisting of spectrometers,fluorescence detectors, colorimeters, light intensity sensors.

In another embodiment, a method is provided for performing an assayusing an analyzer, said method comprising: providing an analyzer havinga processor, a location for holding a plurality or curettes, a locationfor holding a multiplicity of pipette tips, sensors, and a fluidhandling system; introducing reagents into at least some of thecurettes; using the fluid handling system to both (1) transfer reagentsbetween curettes and between curettes and pipette tips and (2) move bothcurettes and pipette tips to sensors to analyze the sample.

It should be understood that any of the foregoing may be configured tohave one or more of the following features. By way of non-limitingexample, one method of the cuvettes are located on a cartridge which isloaded with reagents and thereafter delivered to the analyzer.Optionally, pipette tips are attached to and removed from pipettes whichare part of the fluid handling system. Optionally, pipettes and thepipettes are used to selectively attach to curettes and to move thecurettes within the analyzer.

Other goals and advantages of the invention will be further appreciatedand understood when considered in conjunction with the followingdescription and accompanying drawings. While the following descriptionmay contain specific details describing particular embodiments of theinvention, this should not be construed as limitations to the scope ofthe invention but rather as an exemplification of preferableembodiments. For each aspect of the invention, many variations arepossible as suggested herein that are known to those of ordinary skillin the art. A variety of changes and modifications can be made withinthe scope of the invention without departing from the spirit thereof.

INCORPORATION BY REFERENCE

All publications, patents, and patent applications mentioned in thisspecification are herein incorporated by reference to the same extent asif each individual publication, patent, or patent application wasspecifically and individually indicated to be incorporated by reference.

BRIEF DESCRIPTION OF THE DRAWINGS

The novel features of the invention are set forth with particularity inthe appended claims. A better understanding of the features andadvantages of the present invention will be obtained by reference to thefollowing detailed description that sets forth illustrative embodiments,in which the principles of the invention are used, and the accompanyingdrawings of which:

FIG. 1 shows an example of a system comprising a sample processingdevice and an external controller in accordance with an embodiment ofthe invention.

FIG. 2 shows an example of a sample processing device.

FIG. 3 shows an example of a module having a sample preparation station,assay station, detection station, and a fluid handling system.

FIG. 4 provides an example of a rack supporting a plurality of moduleshaving a vertical arrangement.

FIG. 5 provides an example of a rack supporting a plurality of moduleshaving an array arrangement.

FIG. 6 illustrates a plurality of modules having an alternativearrangement.

FIG. 7 shows an example of a sample processing device having a pluralityof modules.

FIG. 7A shows a non-limiting example of a sample processing devicehaving a plurality of modules.

FIG. 7B shows a non-limiting example of a sample processing devicehaving a plurality of modules.

FIG. 7C shows a non-limiting example of a sample processing devicehaving a plurality of modules.

FIG. 8 shows a plurality of racks supporting one or more modules.

FIG. 9 shows an example of a module with one or more componentscommunicating with a controller.

FIG. 10 shows a system having a plurality of modules mounted in bays(including, e.g., on the racks).

FIG. 11 shows a plurality of plots illustrating a parallel processingroutine.

FIG. 12 shows an exploded view of a positive displacement pipette.

FIG. 13 shows a side view of a positive displacement pipette at a fullaspiration position.

FIG. 14 shows a side view of a positive displacement pipette at a fulldispense position.

FIG. 15 shows an exterior view of an air displacement pipette.

FIG. 16 shows a cross-sectional view of an air displacement pipette.

FIG. 17 shows a close-up of an interface between a pipette tip and anozzle.

FIG. 18 shows an example of an actuation removal mechanism.

FIG. 19A shows a multi-head pipette in accordance with an embodiment ofthe invention.

FIG. 19B shows a side view of a pipette.

FIGS. 20A to C show cross-sectional views of an air displacementpipette. FIG. 20A shows a plunger in a down position and a removalmechanism in a down position; FIG. 20B shows a plunger in anintermediate position and a removal mechanism in an up position; FIG.20C shows a plunger in an up position and a removal mechanism in an upposition.

FIG. 21 shows a plurality of pipettes with removal mechanisms.

FIG. 22 shows an example of a multi-head pipette in accordance with anembodiment of the invention.

FIG. 23 provides an example of a multi-head pipette provided inaccordance with another embodiment of the invention.

FIG. 24 provides an illustration of a vessel that may be used fornucleic acid assays in accordance with an embodiment of the invention.

FIG. 25 illustrates a method for using a vessel in accordance withanother embodiment of the invention.

FIG. 26A provides an illustration of a vessel that may be used forcentrifugation in accordance with an embodiment of the invention.

FIG. 26B provides an illustration of a tip that may be used forcentrifugation in accordance with an embodiment of the invention.

FIG. 27 provides an illustration of a tip that may be used for fluidhandling.

FIG. 28 shows an example of a well.

FIG. 29 illustrates an example of a bulk handling tip in accordance withan embodiment of the invention.

FIG. 30 is an example of an assay tip that may provide colorimetricreadout.

FIG. 31 illustrates an example of a sample tip for processing orfractioning a sample, such as a blood sample.

FIG. 32 is an example of a current reaction tip.

FIG. 33 illustrates an interface between a minitip nozzle and a minitip.

FIG. 34 provides examples of minitips.

FIG. 35 provides an illustration of a microcard and substrate withmicrotips in accordance with an embodiment of the invention.

FIG. 36 shows an example of a centrifuge provided in accordance with anembodiment of the invention.

FIG. 37 provides another example of a centrifuge in accordance with anembodiment of the invention.

FIG. 38 shows an additional example of a centrifuge provided inaccordance with another embodiment of the invention.

FIG. 39 shows a system comprising devices communicating with an externaldevice over a network.

FIG. 40 illustrates a method of processing a sample provided inaccordance with an embodiment of the invention.

FIG. 41A shows an SPI (serial peripheral interface) bridge scheme havingmaster and parallel-series SPI slave bridges. FIG. 41B shows an exampleof an SPI bridge. FIG. 41C shows a module component diagram withinterconnected module pins and various components of a master bridge andslave bridge. FIG. 41D shows slave bridges connected to a master bridge.FIG. 41E shows a device having a plurality of modules mounted on a SPIlink of a communications bus of the device.

FIG. 42 shows an operational matrix of a point of service system.

FIG. 43 is an example of an operational matrix of a point of servicesystem and/or one or more modules of the point of service system.

FIG. 44 shows an operational matrix and a routine matrix.

FIGS. 45A-45C show examples of operational matrices having routines andprocessing states.

FIG. 46 shows an example of a fluid handling apparatus in a retractedposition, provided in accordance with an embodiment of the invention.

FIG. 46A shows a collapsed fluid handling apparatus as previouslydescribed, in a fully retracted position.

FIG. 46B shows a retracted fluid handling apparatus, in a full z-dropposition.

FIG. 47 shows an example of a fluid handling apparatus in an extendedposition in accordance with an embodiment of the invention.

FIG. 48 shows a front view of a fluid handling apparatus.

FIG. 49 shows a side view of a fluid handling apparatus.

FIG. 50 shows another side view of a fluid handling apparatus.

FIG. 51 shows a rear perspective view of a fluid handling apparatus.

FIG. 52 provides an example of a fluid handling apparatus used to carrya sample processing component.

FIG. 53 shows a side view of a fluid handling apparatus useful forcarrying a sample processing component.

FIG. 54 shows an example of a cam-switch arrangement in accordance withan embodiment of the invention. FIG. 54A shows an example of a binarycam at zero position, with the cam rotated zero degrees. FIG. 54B showsan example of a binary cam at position one, with the cam rotated 22.5degrees. FIG. 54C shows an example of a binary cam at position five,with the cam rotated 112.5 degrees. FIG. 54D shows an example of abinary cam at position fifteen, with the cam rotated 337.5 degrees. FIG.54E shows a selection cam mounted with a motor in accordance with anembodiment of the invention.

FIG. 55 shows an example of a fluid handling apparatus using one or morelight source in accordance with an embodiment of the invention. FIG. 55Ashows a plurality of pipette heads. FIG. 55B shows a side cut away viewof a fluid handling apparatus. FIG. 55C shows a close up of a lightsource that may be provided within a fluid handling apparatus. FIG. 55Dshows a close up of a plunger and pipette nozzle. FIG. 55E shows aperspective view of a fluid handling apparatus.

FIG. 56 shows a point of service device having a display, in accordancewith an embodiment of the invention. The display includes a graphicaluser interface (GUI).

FIG. 57 shows a table listing examples of sample preparations.

FIG. 58 shows a table listing examples of possible assays.

FIG. 59 shows an example of a tip interface that includes an example ofa screw-mechanism.

FIG. 60 provides an additional example of a nozzle-tip interface using aclick-fit interface.

FIG. 61 shows an example of an internal screw pick-up interface.

FIG. 62 illustrates an example of an O-ring tip pick-up interface.

FIG. 63 provides an example of an expand/contract smart material tippick-up interface.

FIG. 64 provides an example of an expand/contract elastomer deflectiontip pick-up interface.

FIG. 65 provides an example of a vacuum gripper tip pick-up interface.

FIG. 66 provides an example of a pipette module in accordance with anembodiment of the invention.

FIG. 67A shows an example of modular pipette having a raised shuttle ina full dispense position.

FIG. 67B shows an example of modular pipette having a lowered shuttle ina full dispense position.

FIGS. 67C and 67D show non-limiting examples of pipette configurationsaccording to embodiments described herein.

FIG. 68A provides a top view of an example of a magnetic control.

FIG. 68B provides a side view of the magnetic control.

FIG. 69 provides an example of a cuvette and cuvette carrier.

FIG. 70A shows an example of a carrier (e.g., cuvette), in accordancewith an embodiment of the invention.

FIG. 70B shows additional views of a carrier (e.g., cuvette).

FIG. 71 shows an example of a tip.

FIG. 72 an example of a vial strip.

FIG. 73 shows another example of a vial strip.

FIGS. 74A-74G show non-limiting examples of spectrophotometers accordingto embodiments described herein.

FIGS. 75-76 show non-limiting examples of embodiments of cartridges asdescribed herein.

FIG. 77-78 show non-limiting examples of cartridge covers according toembodiments described herein.

FIG. 79 shows non-limiting examples of absorbant pad assembly accordingto embodiments described herein.

FIG. 80 shows a non-limiting example of sample processing tip accordingto embodiments described herein.

FIGS. 81A and 81B show non-limiting examples of cartridges with thermalconditioning element(s) according to embodiments described herein.

FIGS. 82 to 83 show non-limiting examples of microfluidic cartridgesaccording to embodiments described herein.

FIG. 84 shows a non-limiting example of a cartridge according toembodiments described herein.

FIGS. 85 to 90 show non-limiting examples of thermal conditioningelement(s) according to embodiments described herein.

FIG. 91 shows non-limiting example of a positive displacement tipinterface according to embodiments described herein

FIGS. 92 to 93 show non-limiting examples of an array of sample vesselsaccording to embodiments described herein.

FIGS. 94 to 98 show non-limiting examples of centrifuge vessel imagingconfigurations according to embodiments described herein.

FIGS. 99 to 100 show non-limiting examples of electrochemical sensorconfigurations according to embodiments described herein.

FIG. 101 shows an example of a nucleic acid assay station.

FIG. 102 shows a graph of the relationship between calcium concentrationand absorbance at 570 nm for calcium assays performed on a deviceprovided herein.

FIG. 103 shows a graph of absorbance of multiple measurements over timeof different NADH-containing solutions with a spectrophotometer providedherein.

FIG. 104 shows a graph of the relationship between NADH concentrationand absorbance at 340 nm for measurements performed on spectrophotometerprovided herein and a commercial spectrophotometer.

FIG. 105 shows a graph of the relationship between urea concentrationand absorbance at 630 nm for measurements performed on spectrophotometerprovided herein and a commercial spectrophotometer.

FIGS. 106 to 110 show non-limiting examples of embodiments of modulesaccording to embodiments described herein.

DETAILED DESCRIPTION OF THE INVENTION

While various embodiments of the invention have been shown and describedherein, it will be obvious to those skilled in the art that suchembodiments are provided by way of example only. Numerous variations,changes, and substitutions may occur to those skilled in the art withoutdeparting from the invention. It should be understood that variousalternatives to the embodiments of the invention described herein may beemployed in practicing the invention.

The term “module,” as used herein, refers to a device, component, orapparatus that includes one or more parts or independent units that areconfigured to be part of a larger device or apparatus. In some cases, amodule works independently and independently from another module. Inother cases, a module works in conjunction with other modules (e.g.,modules within modules) to perform one or more tasks, such as assaying abiological sample.

The term “sample handling system,” as used herein, refers to a device orsystem configured to aid in sample imaging, detecting, positioning,repositioning, retention, uptake and deposition. In an example, a robotwith pipetting capability is a sample handling system. In anotherexample, a pipette which may or may not have (other) roboticcapabilities is a sample handing system. A sample handled by a samplehandling system may or may not include fluid. A sampling handling systemmay be capable of transporting a bodily fluid, secretion, or tissue. Asampling handling system may be able to transport one or more substancewithin the device that need not be a sample. For example, the samplehandling system may be able to transport a powder that may react withone or more sample. In some situations, a sample handling system is afluid handling system. The fluid handling system may comprise pumps andvalves of various types or pipettes, which, may comprise but not belimited to a positive displacement pipette, air displacement pipette andsuction-type pipette. The sample handling system may transport a sampleor other substance with aid of a robot as described elsewhere herein.

The term “health care provider,” as used herein, refers to a doctor orother health care professional providing medical treatment and/ormedical advice to a subject. A health care professional may include aperson or entity that is associated with the health care system.Examples of health care professionals may include physicians (includinggeneral practitioners and specialists), surgeons, dentists,audiologists, speech pathologists, physician assistants, nurses,midwives, pharmaconomists/pharmacists, dietitians, therapists,psychologists, chiropractors, clinical officers, physical therapists,phlebotomists, occupational therapists, optometrists, emergency medicaltechnicians, paramedics, medical laboratory technicians, medicalprosthetic technicians, radiographers, social workers, and a widevariety of other human resources trained to provide some type of healthcare service. A health care professional may or may not be certified towrite prescriptions. A health care professional may work in or beaffiliated with hospitals, health care locations and other servicedelivery points, or also in academic training, research andadministration. Some health care professionals may provide care andtreatment services for patients in private or public domiciles,community centers or places of gathering or mobile units. Communityhealth workers may work outside of formal health care institutions.Managers of health care services, medical records and health informationtechnicians and other support workers may also be medical careprofessionals or affiliated with a health care provider. A health careprofessional may be an individual or an institution that providespreventive, curative, promotional or rehabilitative health care servicesto individuals, families, or communities.

In some embodiments, the health care professional may already befamiliar with a subject or have communicated with the subject. Thesubject may be a patient of the health care professional. In someinstances, the health care professional may have prescribed the subjectto undergo a clinical test. The health care professional may haveinstructed or suggested to the subject to undergo a clinical testconducted at the point of service location or by a laboratory. In oneexample, the health care professional may be the subject's primary carephysician. The health care professional may be any type of physician forthe subject (including general practitioners, referred practitioners orthe patient's own physician optionally selected or connected throughtelemedicine services, and/or specialists). The health care professionalmay be a medical care professional.

The term “rack,” as used herein, refers to a frame or enclosure formounting multiple modules. The rack is configured to permit a module tobe fastened to or engaged with the rack. In some situations, variousdimensions of the rack are standardized. In an example, a spacingbetween modules is standardized as multiples of at least about 0.5inches, or 1 inch, or 2 inches, or 3 inches, or 4 inches, or 5 inches,or 6 inches, or 7 inches, or 8 inches, or 9 inches, or 10 inches, or 11inches, or 12 inches.

The term “cells,” as used in the context of biological samples,encompasses samples that are generally of similar sizes to individualcells, including but not limited to vesicles (such as liposomes), cells,virions, and substances bound to small particles such as beads,nanoparticles, or microspheres. Characteristics include, but are notlimited to, size; shape; temporal and dynamic changes such as cellmovement or multiplication; granularity; whether the cell membrane isintact; internal cell contents, including but not limited to, proteincontent, protein modifications, nucleic acid content, nucleic acidmodifications, organelle content, nucleus structure, nucleus content,internal cell structure, contents of internal vesicles, ionconcentrations, and presence of other small molecules such as steroidsor drugs; and cell surface (both cellular membrane and cell wall)markers including proteins, lipids, carbohydrates, and modificationsthereof.

As used herein, “sample” refers to an entire original sample or anyportion thereof, unless the context clearly dictates otherwise.

The invention provides systems and methods for multi-purpose analysis ofa sample or health parameter. The sample may be collected and one ormore sample preparation step, assay step, and/or detection step mayoccur on a device. Various aspects of the invention described herein maybe applied to any of the particular applications, systems, and devicesset forth below. The invention may be applied as a stand alone system ormethod, or as part of an integrated system, such as in a systeminvolving point of service health care. In some embodiments, the systemmay include externally oriented imaging technologies, such as ultrasoundor MRI or be integrated with external peripherals for integrated imagingand other health tests or services. It shall be understood thatdifferent aspects of the invention can be appreciated and practiceindividually, collectively, or in combination with each other.

In accordance with an aspect of the invention, systems for multi-purposeanalysis or analyses and/or sample handling may be provided.

FIG. 1 illustrates an example of a system. A system may comprise one ormore sample processing device 100 that may be configured to receive asample and/or to conduct multi-purpose analysis of one or more sample(s)or types of samples sequentially or simultaneously. Analysis may occurwithin the system. Analysis may or may not occur on the device. A systemmay comprise one, two, three or more sample processing devices. Thesample processing devices may or may not be in communication with oneanother or an external device. Analysis may or may not occur on theexternal device. Analysis may be affected with the aid of a softwareprogram and/or a health care professional. In some instances, theexternal device may be a controller 110.

Systems for multi-purpose analysis may comprise one or more groups ofsample processing devices. Groups of sample processing devices maycomprise one or more device 100. Devices may be grouped according togeography, associated entities, facilities, rooms, routers, hubs, careproviders, or may have any other grouping. Devices within groups may ormay not be in communication with one another. Devices within groups mayor may not be in communication with one or more external devices.

Sample processing devices may comprise one, two or more modules 130.Modules may be removably provided to the devices. Modules may be capableof effecting a sample preparation step, assay step, and/or detectionstep. In some embodiments, each module may be capable of effecting asample preparation step, assay step, and detection step. In someembodiments, one or more modules may be supported by a support structure120, such as a rack. Zero, one, two or more rack(s) may be provided fora device.

Modules may comprise one, two or more components 140 that may be capableof effecting a sample preparation step, assay step, and/or detectionstep. Module components may also include reagents and/or vessels orcontainers that may enable a sample preparation step, assay step, and/ordetection step. Module components may assist with the sample preparationstep, the assay step, and/or detection step. A device may comprise oneor more component that is not provided within a module. In someinstances, a component may be useful for only one of a samplepreparation step, assay step, and/or detection step. Examples ofcomponents are provided in greater detail elsewhere herein. A componentmay have one or more subcomponents.

In some instances, a hierarchy may be provided wherein a systemcomprises one or more groups of devices, a group of devices comprisesone or more device, a device may optionally comprise one or more rackwhich may comprise one or more module, a device may comprise one or moremodule, a module and/or device may comprise one or more components,and/or a component may comprise one or more subcomponents of thecomponent. One or more level of the hierarchy may be optional and neednot be provided in the system. Alternatively, all levels of hierarchydescribed herein may be provided within the system. Any discussionherein applying to one level of hierarchy may also apply to other levelsof hierarchies.

A sample processing device is provided in accordance with an aspect ofthe invention. A sample processing device may comprise one or morecomponents. The sample processing device may be configured to receive asample and/or to conduct one or more sample preparation step, assaystep, and/or detection step. The sample preparation step, assay step,and/or detection step may be automated without requiring humanintervention.

In some embodiments, a system provided herein may be configured asfollows: The system may contain a sample processing device and,optionally an external device. The external device may be, for example,a remote server or cloud-based computing infrastructure. The sampleprocessing device may contain a housing. Within the housing of thedevice, there may be one or more modules. The modules may be supportedby a rack or other support structure. The modules may contain one ormore components or stations. Components and stations of a module mayinclude, for example, assay stations, detection stations, samplepreparation stations, nucleic acid assay stations, cartridges,centrifuges, photodiodes, PMTs, spectrophotometers, optical sensors(e.g. for luminescence, fluorescence, absorbance, or colorimetry),cameras, sample handling systems, fluid handling systems, pipettes,thermal control units, controllers, and cytometers. Components andstations of a module may be removable or insertable into the module. Thecomponents and stations of a module may contain one or moresub-components or other items which may be part of or may be supportedby a component or station. Sub-components may include, for example,assay units, reagent units, tips, vessels, magnets, filters, andheaters. Sub-components of a components or station may be removable orinsertable into the component or station. In addition, the device maycontain one or more additional components which may be part of a module,or which may be elsewhere in the device (e.g. on the housing, rack, orbetween modules) such as a controller, communication unit, power unit,display, sample handing system, fluid handling system, processor,memory, robot, sample manipulation device, detection unit. The system ordevice may have one or more cartridges. The cartridges may be insertableor removable from the device. The cartridges may contain, for examplereagents for performing assays or biological samples. The device mayhave one or more controllers, including one or both of device-level andmodule-level controllers (e.g. where the device level controller isconfigured to direct certain procedures to be performed on certainmodules and where the module level controller is configured to directthe components or stations to execute particular steps for samplepreparation, sample assaying, or sample detection. In an alternative, adevice-level controller may be connected to modules and components ofthe module, to perform both of these functions). The device may have oneor more sample handling system, including both device-level andmodule-level sample handling system (e.g. where the device level samplehandling system is configured to move samples or components betweenmodules and where the module level sample handling system is configuredto move samples or components within a module. In an alternative, adevice level sample handling system may be configured to perform both ofthese functions). The sample processing device may be in two-waycommunication with the external device, such that the sample processingdevice is configured to send information to the external device, andalso to receive information from the external device. The externaldevice may, for example, send protocols to the sample processing device.

In some embodiments, a device may be or comprise a cartridge. Thecartridge may be removable from a large device. Alternatively, thecartridge may be permanently affixed to or integral to the device. Thedevice and/or the cartridge may (both) be components of a disposablesuch as a patch or pill. In some embodiments, an assay station maycomprise a cartridge.

A cartridge may be a universal cartridge that can be configured for thesame selection of tests. Universal cartridges may be dynamicallyprogrammed for certain tests through remote or on-board protocols. Insome cases, a cartridge can have all reagents on board and optionallyserver-side (or local) control through two-way communication systems. Insuch a case, a system using such a disposable cartridge withsubstantially all assay reagents on board the cartridge may not requiretubing, replaceable liquid tanks, or other aspects that demand manualmaintenance, calibration, and compromise quality due to manualintervention and processing steps. Use of a cartridge provided hereincontaining all reagents within the cartridge necessary for performingone or more assays with a system or device provided herein may permitthe device or system to not have any assay reagents or disposablesstored within the device.

Referring now to FIG. 75, one embodiment of a cartridge 9900 will now bedescribed. This embodiment shows that there may be a plurality ofdifferent regions 9920 to 9940 on the cartridge 9900 to providedifferent types of devices, tips, reagents, reaction locations, or thelike. The mix of these elements depends on the types of assays to beperformed using the cartridge 9900. By way of nonlimiting example, thecartridge 9900 may have regions to accommodate one or more samplecontainers, pipette tips, microscopy cuvette, large volume pipette tip,large volume reagent well, large volume strip, cuvette with a lineararray of reaction vessel, round vessels, cap-removal tip, centrifugevessel, centrifuge vessel configured for optical measurement(s), nucleicacid amplification vessels. Any one of the foregoing may be in thedifferent regions 9920 to 9940. Some may arrange the tips and vessels inarrays similar to those of the cartridges shown in commonly assignedU.S. Pat. No. 8,088,593, fully incorporated herein by reference for allpurposes.

By way of non-limiting example, the reagents may also vary in thecartridge and may be selected to include at least those desired toperform at least two or more types of assay panels such as but notlimited to the lipid panel and a chem14 panel or other combination oftwo or more different laboratory testing panels. For example, somecartridges may have reagents, diluents, and/or reaction vessels tosupport at least two different assay types from nucleic acidamplification, general chemistry, immunoassay, or cytometry.

Any one or more of the components of the cartridge may be accessible bya sample handling system of the system. The different zones in thecartridge may be configured to match the pitch of the pipette heads usedin the system. Optionally, some zones are configured to be at pitchesthat are multiples of or fractions of the pitch of the pipette heads.For example, some components of the cartridge are at ⅓× of the pitch,others at ½× of the pipette pitch, others at a 1× pitch, others at a 2×pitch, while still others at a 4× pitch.

Referring still to FIG. 75, it should be understood that there may becomponents located at one plane of the cartridge while other are locatedat lower or higher planes. For example, some components may be locatedbelow a cuvette or other component. Thus, once the upper component isremoved, the lower components become accessible. This multi-layerapproach provides for greater packing density in terms of components ona cartridge. There may also be locating features on the cartridge 9900such as but not limited to rail 9834 that is configured to engagematching slot on the cartridge receiving location in the system. Thecartridge may also have registration features (physical, optical, or thelike) that allow the system to accurately engage components of thecartridge once the cartridge is recognized by the system. By way ofnon-limiting example, although components may be removed from thecartridge 9900 during assay processing, it is understood that someembodiments may permit the return of all components back to thecartridge for unified disposal. Optionally, in some embodiments of thesystem may have disposal areas, containers, chutes, or the like todiscard those components of the cartridge not returned to the cartridgeprior to ejecting the cartridge from the system. In some embodiments,these areas may be dedicated areas of the system for receiving waste.

Referring now to FIG. 76, another embodiment of cartridge 9901 will nowbe described. This one uses a reduced height cartridge 9901 wherein thesidewalls have a reduced vertical height. The provides for less materialuse for the disposable and brings the reaction vessels and/or reagents.

Referring now to FIG. 77, yet another feature of at least somecartridges will now be described. FIG. 77 shows a side view of acartridge 9900 with a lid 9970, wherein the lid 9970 is removable uponinsertion of the cartridge 9900 into the system and will re-engage thecover when the cartridge 9900 is removed from the system. Such featuresmay be advantageous for increasing the security and protection of thecomponents of the cartridge (e.g. to prevent tampering or inadvertentintroduction of external matter). As seen in FIG. 77, there is anengagement feature 9972 such as but not limited to snap that engages alocking feature 9974 in the body of the cartridge 9900. A releasemechanism 9976 such as but not limited to a pin can be inserted into anopening where it can contact the locking feature 9974 and move it to arelease position. This allows one end of the lid 9970 to be disengagedautomatically when the cartridge 9900 is inserted into system.Optionally, the release mechanism 9976 may have pins that actuate sothat the release of the lid 9970 is based on when the system actuates tounlock the locking feature 9974. In one non-limiting example, a springmechanism 9980 such as but not limited to a torsional spring canautomatically lift open the lid 9970 as indicated by arrow 9982 afterthe locking mechanism 9974 is disengaged. When ejecting the cartridge9900, the motion of the cartridge 9900 out of the device will cause thelid 9970 in the open position to engage a horizontally or otherwisemounted closure device 9984 (shown in phantom) that will move the lid9970 to a closed position due the motion of the cartridge 9900 asindicated by arrow 9986 as it passes under the device 9984. In thepresent embodiment, the spring mechanism 9980 is engaged to thecartridge 9900 through openings 9978 (see FIG. 75).

FIG. 78 shows a perspective view of one embodiment of the lid 9970 thatengages over a cartridge 9900. This lid may be configured to retain allof the various components of the cartridge 9900 inside the cartridgewhen the cartridge is not in the system. The use of dual engagementfeatures 9972 more securely holds the lid 9970 to the cartridge andmakes it more difficult for a user to accidentally open the lid 9970 asit uses two or more points of engagement with the locking mechanism ofthe cartridge. As seen in FIG. AC, there is also a cut-out portion 9988that allow for the sample containers to be placed into the cartridge9900 before the cartridge 9900 is loaded into the system. In onenon-limiting example, this can simplify use of the cartridge as this isonly allows the sample container(s) to be placed in one location in thecartridge 9900, thus making the user interaction with the cartridge forloading sample much less variable or subject to error. The lid 9970 canalso be opaque to prevent the user from being distracted by vessels andelements in the cartridge, instead focusing the user's attention to theonly available open slot, which in the current embodiment is reservedfor the sample container(s) which can only be inserted in a particularorientation due to the keyed shape of the opening.

Referring now to FIG. 79, it should be understood that the cartridge9900 may also contain an absorbent pad assembly 10000 that is used toremove excess fluid from the various tips, vessels, or other elements.In one embodiment, the absorbent pad assembly 10000 has a multi-layerconfiguration comprising a spacer 10002, the absorbent pad 10004, and anadhesive layer 10006. Some embodiments may or may not have the spacerlayer 10002 which may be made of material such as but not limited toacrylic or other similar material. The shape of the openings in thespacer 10002 is sized to allow for features such as but not limited topipettes tip to enter spacer layer 10002 to clean the tip for excessfluid without contaminating the absorbent pad 10004 for adjacentopenings. Optionally, it should be understood that the absorbentmaterial 10004 may also be used alone or with adhesive or other materialto cover certain reagent or other zones such that a tip would penetratethrough the absorbent material 10004 in order to reach the reagentbelow. This would provide for removal of excess fluid on the outside ofthe tips on insertion and/or withdrawal of the tip, and may aid in thereduction of cross-reactivity. In one embodiment, this may be like aburst-able membrane of the absorbent material. Some embodiments may usetips that are linear and not conical in shape at the distal portion sothat contact with the absorbent material is not lost due to variation intip diameter, resulting in a less than thorough wiping of fluid from anoutside portion of the tip.

In some embodiments, tips may be configured such that they do not retainexcess fluid on the outside of the tip, and are not used with anabsorbent pad.

Referring now to FIG. 80, it should be understood that the cartridge mayalso include various types of specialized tips or elements for specificfunctions. By way of non-limiting example as seen in FIG. 80, a samplepreparation tip 10050 will now be described. In this embodiment of asample preparation tip, the plunger 10052 of the tip 10050 interfaceswith a single minitip nozzle at opening 10054; the pipette nozzle can beset to “pull” to produce a vacuum that allows the plunger to stay on thenozzle more securely. In the present embodiment, the barrel part 10056of the sample preparation tip 10050 interfaces with two minitip nozzlesof the pipette at cavities 10058 and 10060. In this manner, the pipettesystem uses multiple heads with nozzles thereon to both move thehardware of the tip 10050 and to aspirate using the plunger 10052.

In the present embodiment, the tip 10050 may include a resin portion10070 that may be bound above and below by frits 10072 and 10074. Fritmaterial may be compatible with sample purification chemistry and notleach any carryover inhibition into the downstream assay. Optionally,frit material should not bind to the biomolecule of interest, or must bechemically treated or surface passivated to prevent such. Optionally,frit material may be porous with an appropriate pore such that the resinremains within the confines of its cavity. Optionally, frit must besized such that the interference fit between the barrel and the frit isenough to hold it in place against typical operating fluid pressures. Byway of non-limiting example, the resin portion 10070 may be chosen suchthat it binds optimally with the biomolecule of choice, which caninclude but is not limited to bare and chemically modified versions ofsilica, zirconia, polystyrene or magnetic beads.

In one embodiment, the method for using the tip 100050 may involve theaspiration of lysed unpurified sample mixed with binding buffer throughthe resin 10070. In such an example, DNA or biomolecule of choice willbind to the resin 10070 in the appropriate salt conditions and remainingfluid is dispensed into waste. The method may involve aspiration of washbuffers to clean the bound sample and dispense fluid into waste vessel.This may be repeated multiple times as desired to obtain a clean sample.The method may further include aspiration and dispense of heated air inorder to dry to resin to remove residual solvents and any carryoverinhibition that may interfere with the downstream assay. Optionally, thetip 10050 may be used for aspiration of elution buffer to remove thebound molecule of interest, and may allow the elution buffer tocompletely saturate the resin before dispensing into an appropriatecollection vessel.

In some embodiments, a pipette tip may contain a septa, such that thereis a seal between the sample intake portion of a pipette tip, and thepath of an actuation mechanism of the pipette (e.g. the piston block).

In some embodiments, a pipette nozzle and pipette tip may have threads,such that the pipette tip may be threaded onto the tip (e.g. byrotation). The nozzle may rotate to thread the tip onto the nozzle, orthe tip may rotate. The tip may be “locked” in place on the nozzle uponthreading the tip onto the pipette nozzle. The tip may be “unlocked” byrotating the nozzle or the tip in the opposite direction as used forloading the tip onto the nozzle.

Referring now to FIG. 81, in some embodiments, the cartridge 9800contains at least one thermal device 9802 such as a chemical reactionpack for generating heat locally to enhance kinetics and/or for heatinga mixture. The chemical reaction pack may contain chemicals such assodium acetate or calcium chloride. This may be particularly desirablein situations where the cartridge 9800, prior to use, is stored in arefrigerated condition such as but not limited to the 0° C. to 8° C.range for days to weeks. Optionally, the temperature range during coldstorage may be in the range of about −20° C. to 8° C., optionally −10°C. to 5° C., optionally −5° C. to 5° C., or optionally 2° C. to 8° C. Inone non-limiting example, the thermal pack 9802 is in a refrigeratedcondition for at least one month. In an implementation, sodium acetateis used in the chemical in the chemical reaction thermal pack 9802.Sodium acetate trihydrate crystals melt at 58.4° C., dissolving inwater. When they are heated to around 100° C., and subsequently allowedto cool, the aqueous solution becomes supersaturated. This solution iscapable of cooling to room temperature without forming crystals. Whenthe supersaturated solution is disrupted, crystals are formed. Thebond-forming process of crystallization is exothermic. The latent heatof fusion is about 264-289 kJ/kg. The crystallization event can betriggered by clicking on a metal disc, creating a nucleation centerwhich causes the solution to crystallize into solid sodium acetatetrihydrate again. This can be triggered by the pipette in the system orother actuator in the device. Alternatively, a tip/needle on the pipettewith sodium acetate crystal on its surface can puncture the sodiumacetate foil seal. This will also trigger crystallization. It should beunderstood that other exothermic reactions can be used instead of sodiumacetate and these other reactions are not excluded. One non-limitingexample is to use magnesium/iron alloy in a porous matrix formed frompolymeric powders with sodium chloride incorporated. The reaction isstarted by the addition of water. The water dissolves the sodiumchloride into an electrolyte solution causing magnesium and iron tofunction as an anode and cathode, respectively. Optionally, anexothermic oxidation-reduction reaction between the magnesium-iron alloyand water can be used to produce magnesium hydroxide, hydrogen gas andheat. Optionally, a fan or other flow generating device on the systemcan be used to provide convective flow. The fan can be placed to blowair to the underside of the cartridge, along the sides, or optionallyover the tops of the cartridge.

It should be understood that some cartridges 9800 may have more than oneheater. As seen in FIGS. 81A and 81B, a second thermal device 9804 canalso be a part of the cartridge 9800. In some embodiments, the heaters9802 and 9804 are sized and located to thermally control temperature forcertain areas of the cartridge 9800, particularly those vessels, wells,or other features that contain materials that are sensitive totemperature or provide more consistent or accurate results when they areused in certain temperature ranges. As seen in FIGS. 81A and 81B, theheaters 9802 and 9804 are positioned to thermally condition (heat orcool) those locations in the cartridge. In some embodiments, the heaters9802 and 9804 are positioned to thermally condition particular reagentsin a cartridge. It should also be understood that thermally conductivematerial such as but not limited to aluminum, copper, or the like, mayalso be incorporated into the cartridge to preferentially thermallycondition certain areas of the cartridge. In one non-limiting example,the thermally conductive materials 9806 and 9808 can be made of amaterial different from that of the cartridge and be shaped toaccommodate, contour, or otherwise be in contact with or near certainpipette tips, reagent wells, diluent wells, or the like. In someembodiments, the thermally conductive material may be used to conditionthose areas that are spaced apart from the thermal devices to morereadily propagate thermal conditioning to other areas of the cartridge.Optionally, the thermally conductive material is located only attargeted areas over the thermal packs and designed to only thermallycondition some but not other areas of the cartridge. Optionally, someembodiments may integrate thermally conductive materials such as but notlimited to metal beads or other thermally conductive materials into thepolymeric or other material used to form the cartridge. The cartridgecan have isolated regions with temperature control (e.g. a region withhigh temperature for nucleic acid tests), without affecting other partsof the cartridge/device.

Referring now to FIG. 82, in another embodiment, the cartridge receivinglocation 9830 with rails 9832 is configured to receive a cartridgecomprising a microfluidic cartridge 9810. This passive flow cartridge9810 may have one more sample deposit locations 9812. By way ofnon-limiting example, this cartridge 9810 may be a microfluidiccartridge as described in U.S. Pat. Nos. 8,007,999 and 7,888,125, bothfully incorporated herein by reference for all purposes. The passiveflow cartridge 9810 may also have one or more rails that engage at leastone slot 9832 of the cartridge receiving location. The cartridgereceiving location 9830 may also have one more signal interfacelocations on the cartridge such as but not limited to electricalconnectors or optical connectors so that electrodes, fiberoptics, orother elements in the cartridge can communicate with correspondingequipment in the system that can read signals from elements in thecartridge 9810.

It should be understood that a pipette may be used to load sample intothe cartridge 9810. Optionally, the passive flow cartridge 9810 may alsobe integrated for use with the pipette to transport sample from certainports in the cartridge 9810 to other ports on the sample cartridge, toother cartridges, or to other types of sample vessels. After thecompletion, the cartridge may be unloaded from the cartridge receivinglocation 9830 as indicated by arrow 9819.

Referring now to FIG. 83, in a still further embodiment, the cartridgereceiving location 9830 with rails 9832 is configured to receive acartridge comprising a microfluidic portion 9822. In this non-limitingexample, the microfluidic portion 9822 is mounted on a larger cartridge9824 that can have various reagent region(s) 9826 and sample vesselregion(s) 9828. Some embodiments may also have a cartridge with samplevessel holding location 9938 that transports the sample fluid in gastight containers until they are ready for analysis when loaded into thedevice. In one non-limiting example, the sample being aliquoted intomicrofluidic portion 9822 may be pre-treated by material in the samplevessel. In some embodiments, the microfluidic portion 9822 can be movedto location separate from the cartridge 9824 so that the processing onthe microfluidic portion 9822 can occur simultaneously with other sampleprocessing that may occur on the cartridge 9824. Optionally, the systemmay have the microfluidic portion 9822 moved so that other reagents,diluents, tips, or vessels that, in the present embodiment, are housedbelow the microfluidic portion 9822, become accessible for use.Optionally, the microfluidic portion 9822 may be returned to thecartridge 9824 after use. The entire cartridge 9824 may use a cover 9970(not shown) to provide an enclosed unit for improved cartridge handlingwhen not in use in the system.

Referring now to FIG. 84, another embodiment of a cartridge receivinglocation 9830 will now be described. This embodiment shows a pluralityof detector locations 9841 on a cartridge 9842. A pipette 9844 can beused to transport sample to one or more the detector locations 9841. Inone non-limiting example, movement of sample from one detector locations9841 to another, or optionally, from a sample vessel to one or more ofthe detector locations 9841 can be by way of the pipette 9844.

In one non-limiting example, the measurement of the sample at thedetector locations 9841 can be by way of a sensing electrode used in oneof two manners. First, the change can be detected with respect to theexposed reference capacitor. In this embodiment, the reference electrodeis exposed to the same solution as the sensing electrode. Optionally, aprobe is designed to have similar electrical characteristics as theaffinity probed but not to bind to a target in the solution in attachedto the reference electrode. A change in integrated charge is measured asbinding occurs on the sensing electrode (or affinity probe attachedthereon) whose electrical characteristics change, but not on thereference electrode whose electrical characteristic remain the same.Second, two measurements of the same electrode, before and after theanalyte binds, can be compared to establish the change in integratedcharge resulting from binding. In this case, the same electrode at aprevious time provides the reference. The device may operate indifferential detection mode, in which both reference and sense electrodehave attached affinity probes (of different affinity) to reject commonmode noise contributed by the matrix or other noise sources.

In an alternative configuration, the reference electrode can beconfigured so that the sensing electrode takes direct capacitancemeasurements (non-differential). In this configuration, the referenceelectrode can be covered with a small dielectric substance such as epoxyor the device passivation or left exposed to air. The signal from theelectrode can then be compared to an open circuit which establishes anabsolute reference for measurement but may be more susceptible to noise.Such an embodiment uses the device in an absolute detection mode, inwhich the reference is an unexposed (or exposed to a fix environmentsuch as air) fixed capacitor.

Referring now to FIGS. 85 to 88, it should be understood that in someembodiments the thermal device is not integrated into a part of adisposable such as cartridge 9800 but is instead a non-disposable thatis part of the hardware of the system. The thermal device may be athermal control unit. FIG. 85 shows one embodiment of a cartridge 9820that is received into an assay station receiving location 9830 of thesystem. In some embodiments, an assay station receiving location may bea tray. In this non-limiting example, the assay station receivinglocation 9830 has slots 9832 that are shaped to receive rails 9834 onthe cartridge 9820. The cartridge 9820 is inserted into the assaystation receiving location 9830 until the cartridge 9820 engages a stop9836. It should be understood that the regions in FIGS. 85-88 andoptionally in other cartridges described herein, the region may containa plurality of wells, tips or the like such as shown in the cartridgesof U.S. Pat. No. 8,088,593 fully incorporated herein by reference forall purposes.

Referring now to FIG. 86 which shows an underside view of the assaystation receiving location 9830 which shows that there may be convectiveflow devices 9840 positioned on the assay station receiving location9830 to facilitate flow in the underside of the cartridge 9820 when itis in the desired location on the assay station receiving location 9830.Although FIG. 86 shows the devices 9840 in only one location, it shouldbe understood that devices 9840 may also be located at one or more otherlocations to access other areas of the cartridge 9820. Some embodimentsmay configure at least one of the convective devices 9840 to be pullingin air while at least one other convective device 9840 is pushing airout of the cartridge. There may be features such as but not limited tovanes, fins, rods, tubes, or the like to guide air flow in the undersideor other areas of the cartridge 9820.

Referring now to FIG. 87, a cross-sectional view is shown of thecartridge 9820 on the assay station receiving location 9830 that ispositioned over the convective flow device 9840. FIG. 0 further showsthat there is thermal device 9850 that is a non-disposable that remainspart of the system and is not disposed with the cartridge.Alternatively, some embodiments may integrate the thermal device 9850into the cartridge, in which case the thermal device 9850 is part of thedisposable. As seen in FIG. 87, the thermal device 9850 is at a firstlocation spaced apart from the targeted materials 9852 to be thermallyconditioned in the cartridge 9820. Referring still to FIG. 87, in someembodiments, the underside of the cartridge is substantially enclosedexcept for perhaps a hatch, door, or cover that allows for access to theunderside of the cartridge 9820.

Referring now to FIG. 88, this illustration shows that the thermaldevice 9850 can be moved from the first location to a second location tomore directly contact the areas and/or components of the cartridge 9820to be thermally conditioned. As seen in FIG. 88, the thermal device 9850can have shapes such as but not limited to cavities, openings, or thelike that are contoured to engage surfaces of the areas and/orcomponents of the cartridge 9820 to be thermally conditioned. It shouldbe understood that the thermal device 9850 can use various thermalelements to heat or cool the portions that engage features of thecartridge or cartridge components. In one non-limiting example, thethermal device 9850 may use heating rods 9852 in the device 9850. Thesemay cause thermal conditioning through electro-resistive heating or thelike. Thermal transfer may occur from corresponding cavities inheater-block into each round-vessel bottom-stem through narrow air-gap.The convective flow device 9840 may assist in accelerating the thermalconditioning. Optionally, some embodiments may use the convective flowdevice 9840 to bring steady state condition to the cartridge soonerafter an initial thermal conditioning phase. By way of non-limitingexample, a pre-heated heater block may be the thermal device 9850 thatengages with refrigerated (e.g. 4° C.) cartridge-round-vessels in thecartridge 9820, followed by rapid heating from thermal device 9850,followed by fan-cooling by convective flow device 9840, which then leadsto controllable operating temperature in vessels within about 180seconds.

After thermally conditioning is completed or to provide better accessfor the convective flow device 9840, the thermal device 9850 optionallyreturns to a location where it does not interfere with the insertionand/or removal of the cartridge 9820 from the assay station receivinglocation 9830, such as but not limited to residing in recess 9858.

Referring now to FIGS. 89 and 90, yet another thermal controlconfiguration will now be described. As seen in FIG. 89, one embodimentshows that the support structure of a module 9870 can be thermallycontrolled. In some embodiments, the support structure of a module maybe a chassis. The support structure of a module 9870, which can have aplurality of components mounted thereon (not shown for ease ofillustration), is then used to provide thermal conditioning to multiplecomponents mounted on the chassis 9870. The support structure of amodule 9870 may have a thermal base plate 9872. The thermal base plate9872 may create a uniform thermal condition for the entire base plate9872 or a portion thereof. By way non-limiting example, the thermalconditioning may be through electroresistive elements embedded in or ona thermally conductive material used for the base plate.

Optionally as seen in FIG. 90, another embodiment may use a supportstructure of a module 9880 that has a non-uniform thermal base plate9882 that selectively thermally conditions one or more location in thebase plate. This can be designed for use with thermally conductive,thermally neutral, or thermally insulating material for the base plate.This allows for creating different thermal zones, depending on thedesired thermal profile for the various operating conditions ofcomponents mounted on the support structure of a module 9880. By way ofnon-limiting example, some embodiments may have a heated location underthe assay station receiving location on the support structure of amodule 9880. When a system uses multiple chassis on rack or othermultiple chassis systems, some embodiment may use only those chassiswith the thermal base plate. Optionally, some embodiments may use a mixof those chassis with or without thermal base plates.

In some embodiments, a both a disposable such as a cartridge and thehardware of the system contain a thermal device. In some embodiments, acartridge is not thermally conditioned prior to or during use.

Optionally, the cartridge can also transform into differentconfigurations based on external or internal stimuli. The stimuli can besensed via sensors on the cartridge body, or be part of the cartridge.More commonplace sensors such as RFID tags can also be part of thecartridge. The cartridge can be equipped with biometric sensors if, forexample, the sample collection and analysis are done in two separatelocations (e.g. for patients in intensive care, samples are collectedfrom the patient and then transferred to the device for analysis). Thisallows linking a patient sample to the cartridge, thereby preventingerrors. The cartridge could have electric and/or fluidic interconnectsto transfer signals and/or fluids between different vessels, tips, etc.on the cartridge. The cartridge can also comprise detectors and/orsensors.

Intelligent cartridge design with feedback, self learning, and sensingmechanisms enables a compact form factor with point of service utility,waste reduction, and higher efficiencies.

In one embodiment, a separate external robotics system may be availableon site to assemble new cartridges in real time as they are needed.Alternatively, this capability could be part of the device or cartridge.Individual cartridge components for running assays may include but arenot limited to sealed vessels with reagents, as well as tips and vesselsfor mixing and optical or non-optical measurements. All or some of thesecomponents can be added to a cartridge body in realtime by an automatedrobotic system. The desired components for each assay can be loadedindividually onto a cartridge, or be pre-packaged into a mini-cartridge.This mini-cartridge can then be added to the larger cartridge which isinserted into the device. One or more assay units, reagent units, tips,vessels or other components can be added to a cartridge in real time.Cartridges may have no components pre-loaded onto them, or may have somecomponents preloaded. Additional components can be added to a cartridgein real time based on a patient order. The position of the componentsadded to a cartridge are predetermined and/or saved so that the deviceprotocol can properly execute the assay steps in the device. The devicemay also configure the cartridge in real time if the assay cartridgecomponents are available to the device. For example, tips and othercartridge components can be loaded into the device, and loaded intocartridges in real time given the patient order to the run at that time.

FIG. 2 shows an example of a device 200. A device may have a samplecollection unit 210. The device may include one or more supportstructure 220, which may support one or more module 230 a, 230 b. Thedevice may include a housing 240, which may support or contain the restof the device. A device may also include a controller 250, display 260,power unit 270, and communication unit 280. The device may be capable ofcommunicating with an external device 290 through the communicationunit. The device may have a processor and/or memory that may be capableof effecting one or more steps or providing instructions for one or moresteps to be performed by the device, and/or the processor and/or memorymay be capable of storing one or more instructions.

Sample Collection

A device may comprise a sample collection unit. The sample collectionunit may be configured to receive a sample from a subject. The samplecollection unit may be configured to receive the sample directly fromthe subject or may be configured to receive a sample indirectly that hasbeen collected from the subject.

One or more collection mechanisms may be used in the collection of asample from a subject. A collection mechanism may use one or moreprinciple in collecting the sample. For example, a sample collectionmechanism may use gravity, capillary action, surface tension,aspiration, vacuum force, pressure differential, density differential,thermal differential, or any other mechanism in collecting the sample,or a combination thereof.

A bodily fluid may be drawn from a subject and provided to a device in avariety of ways, including but not limited to, fingerstick, lancing,injection, pumping, swabbing, pipetting, breathing, and/or any othertechnique described elsewhere herein. The bodily fluid may be providedusing a bodily fluid collector. A bodily fluid collector may include alancet, capillary, tube, pipette, syringe, needle, microneedle, pump,laser, porous membrane or any other collector described elsewhereherein. The bodily fluid collector may be integrated into a cartridge oronto the device, such as through the inclusion of a lancet and/orcapillary on the cartridge body or vessel(s) or through a pipette thatcan aspirate a biological sample from the patient directly. Thecollector may be manipulated by a human or by automation, eitherdirectly or remotely. One means of accomplishing automation or remotehuman manipulation may be through the incorporation of a camera or othersensing device onto the collector itself or the device or cartridge orany component thereof and using the sensing device to guide the samplecollection.

In one embodiment, a lancet punctures the skin of a subject and draws asample using, for example, gravity, capillary action, aspiration,pressure differential and/or vacuum force. The lancet, or any otherbodily fluid collector, may be part of the device, part of a cartridgeof the device, part of a system, or a stand alone component. In anotherembodiment, a laser may be used to puncture the skin or sever a tissuesample from a patient. The laser may also be used to anesthetize thesample collection site. In another embodiment, a sensor may measureoptically through the skin without invasively obtaining a sample. Insome embodiments, a patch may comprise a plurality of microneedles,which may puncture the skin of a subject. Where needed, the lancet, thepatch, or any other bodily fluid collector may be activated by a varietyof mechanical, electrical, electromechanical, or any other knownactivation mechanism or any combination of such methods.

In some instances, a bodily fluid collector may be a piercing devicethat may be provided on a disposable or that may be disposable. Thepiercing device may be used to convey a sample or information about thesample to a non-disposable device that may process the sample.Alternatively, the disposable piercing device itself may process and/oranalyze the sample.

In one example, a subject's finger (or other portion of the subject'sbody) may be punctured to yield a bodily fluid. The bodily fluid may becollected using a capillary tube, pipette, swab, drop, or any othermechanism known in the art. The capillary tube or pipette may beseparate from the device and/or a cartridge of the device that may beinserted within or attached to a device, or may be a part of a deviceand/or cartridge. In another embodiment where no active mechanism(beyond the body) is required, a subject can simply provide a bodilyfluid to the device and/or cartridge, as for example, could occur with asaliva sample or a finger-stick sample.

A bodily fluid may be drawn from a subject and provided to a device in avariety of ways, including but not limited to, fingerstick, lancing,injection, and/or pipetting. The bodily fluid may be collected usingvenous or non-venous methods. The bodily fluid may be provided using abodily fluid collector. A bodily fluid collector may include a lancet,capillary, tube, pipette, syringe, venous draw, or any other collectordescribed elsewhere herein. In one embodiment, a lancet punctures theskin and draws a sample using, for example, gravity, capillary action,aspiration, or vacuum force. The lancet may be part of the readerdevice, part of the cartridge, part of a system, or a stand alonecomponent, which can be disposable. Where needed, the lancet may beactivated by a variety of mechanical, electrical, electromechanical, orany other known activation mechanism or any combination of such methods.In one example, a subject's finger (or other portion of the subject'sbody) may be punctured to yield a bodily fluid. Examples of otherportions of the subject's body may include, but is not limited to, thesubject's hand, wrist, arm, torso, leg, foot, ear, or neck. The bodilyfluid may be collected using a capillary tube, pipette, or any othermechanism known in the art. The capillary tube or pipette may beseparate from the device and/or cartridge, or may be a part of a deviceand/or cartridge or vessel. In another embodiment where no activemechanism is required, a subject can simply provide a bodily fluid tothe device and/or cartridge, as for example, can occur with a salivasample. The collected fluid can be placed within the device. A bodilyfluid collector may be attached to the device, removably attachable tothe device, or may be provided separately from the device.

In some embodiments, a sample may be provided directly to the device, ormay use an additional vessel or component that may be used as a conduitor means for providing a sample to a device. In one example, feces maybe swabbed onto a cartridge or may be provided to a vessel on acartridge. In another example a urine cup may snap out from a cartridgeof a device, a device, or a peripheral to a device. Alternatively, asmall vessel may be pushed out, snapped out, and/or twisted out of acartridge of a device or a peripheral to a cartridge. Urine may beprovided directly to the small vessel or from a urine cup. In anotherexample, a nasal swab may be inserted into a cartridge. A cartridge mayinclude buffers that may interact with the nasal swab. In someinstances, a cartridge may include one or more tanks or reservoirs withone or more reagents, diluents, wash, buffers, or any other solutions ormaterials. A tissue sample may be placed on a slide that may be embeddedwithin a cartridge to process the sample. In some instances, a tissuesample may be provided to a cartridge through any mechanism (e.g.,opening, tray), and a slide may be automatically prepared within thecartridge. A fluid sample may be provided to a cartridge, and thecartridge may optionally be prepared as a slide within the cartridge.Any description of providing a sample to a cartridge or a vessel thereinmay also be applied to providing the sample directly to the devicewithout requiring a cartridge. Any steps described herein as beingperformed by the cartridge may be performed by the device withoutrequiring a cartridge.

A vessel for sample collection can be configured to obtain samples froma broad range of different biological, environmental, and any othermatrices. The vessel can be configured to receive a sample directly froma body part such as a finger or an arm by touching the body part to thevessel. Samples may also be introduced through sample transfer deviceswhich may optionally be designed for single-step processing intransferring a sample into a vessel or cartridge or into the device.Collection vessels may be designed and customized for each differentsample matrix that is processed, such as urine, feces, or blood. Forexample, a sealed vessel may twist off of or pop out of a traditionalurine cup so that it can be placed directly in a cartridge without theneed for pipetting a sample. A vessel for sample collection can beconfigured to obtain blood from a fingerstick (or other puncture site).The collection vessel may be configured with one or more entry portseach connected to one or more segregated chambers. The collection vesselmay be configured with only a single entry port connected to one of moresegregated chambers. The collected sample may flow into the chambers viacapillary action. Each segregated chamber may contain one or morereagents. Each segregated chamber may contain different reagents fromthe other chambers. Reagents in the chambers may be coated on thechamber walls. The reagents may be deposited in certain areas of thechambers, and/or in a graded fashion to control reagent mixing anddistribution in the sample. Chambers may contain anticoagulants (forexample, lithium-heparin, EDTA (ethylenediaminetetraacetic acid),citrate). The chambers may be arranged such that mixing of the sampleamong the various chambers does not occur. The chambers may be arrangedsuch that a defined amount of mixing occurs among the various chambers.Each chamber may be of the same or different size and/or volume. Thechambers can be configured to fill at the same or different rates withthe sample. The chambers may be connected to the entry port via anopening or port that may have a valve. Such a valve may be configured topermit fluid to flow in one or two directions. The valve may be passiveor active. The sample collection vessel may be clear or opaque incertain regions. The sample collection vessel may be configured to haveone or more opaque regions to allow automated and/or manual assessmentof the sample collection process. The sample in each chamber may beextracted by the device by a sample handling system fitted with a tip orvessel to interface with the sample collection vessel. The sample ineach chamber may be forced out of the chamber by a plunger. The samplesmay be extracted or expelled from each chamber individually orsimultaneously.

A sample may be collected from an environment or any other source. Insome instances, the sample is not collected from a subject. Examples ofsamples may include fluids (such as liquids, gas, gels), solid, orsemi-solid materials that may be tested. In one scenario, a food productmay be tested to determine whether the food is safe to eat. In anotherscenario, an environmental sample (e.g., water sample, soil sample, airsample) may be tested to determine whether there are any contaminants ortoxins. Such samples can be collected using any mechanism, includingthose described elsewhere herein. Alternatively, such samples can beprovided directly to the device, cartridge or to a vessel.

The collected fluid can be placed within the device. In some instances,the collected fluid is placed within a cartridge of the device. Thecollected fluid can be placed in any other region of the device. Thedevice may be configured to receive the sample, whether it be directlyfrom a subject, from a bodily fluid collector, or from any othermechanism. A sample collection unit of the device may be configured toreceive the sample.

A bodily fluid collector may be attached to the device, removablyattachable to the device, or may be provided separately from the device.In some instances, the bodily fluid collector is integral to the device.The bodily fluid collector can be attached to or removably attached toany portion of the device. The bodily fluid collector may be in fluidcommunication with, or brought into fluid communication with a samplecollection unit of the device.

A cartridge may be inserted into the sample processing device orotherwise interfaced with the device. The cartridge may be attached tothe device. The cartridge may be removed from the device. In oneexample, a sample may be provided to a sample collection unit of thecartridge. A cartridge may brought to a selected temperature beforebeing inserted into the device (e.g. to 4 C, room temperature, 37 C, 40C, 45 C, 50 C, 60 C, 70 C, 80 C, 90 C, etc.). The sample may or may notbe provided to the sample collection unit via a bodily fluid collector.A bodily fluid collector may be attached to the cartridge, removablyattachable to the cartridge, or may be provided separately from thecartridge. The bodily fluid collector may or may not be integral to thesample collection unit. The cartridge may then be inserted into thedevice. Alternatively, the sample may be provided directly to thedevice, which may or may not use the cartridge. The cartridge maycomprise one or more reagents, which may be used in the operation of thedevice. The reagents may be self-contained within the cartridge.Reagents may be provided to a device through a cartridge withoutrequiring reagents to be pumped into the device through tubes and/ortanks of buffer. Alternatively, one or more reagents may already beprovided onboard the device. The cartridge may comprise a shell andinsertable tubes, vessels, or tips. The cartridge may contain, forexample, assay units, reagent units, processing units, or cuvettes (forexample, cytometry cuvettes). Vessels or tips may be used to storereagents required to run tests. Some vessels or tips may be preloadedonto cartridges. Other vessels or tips may be stored within the device,possibly in a cooled environment as required. At the time of testing,the device can assemble the on-board stored vessels or tips with aparticular cartridge as needed by use of a robotic system within thedevice.

In some embodiments, a cartridge contains microfluidics channels. Assaysmay be performed or detected within microfluidics channels of acartridge. Microfluidics channels of a cartridge have openings tointerface with, for example, tip, such that samples may be loaded intoor removed from the channel. In some embodiments, samples and reagentsmay be mixed in a vessel, and then transferred to a microfluidicschannel of a cartridge. Alternatively, samples and reagents may be mixedwithin a microfluidics channel of a cartridge.

In some embodiments, a cartridge contains chips for electronicmicrofluidics applications. Small volumes of liquids may be applied tosuch chips, and assay may be performed on the chips. Liquids may be, forexample, spotted or pipetted onto the chips, and moved, for example bycharge.

In some embodiments, a cartridge contains one or more assay units,reagent units, or other vessels containing, for example, antibodies,nucleic acid probes, buffers, chromogens, chemiluminescent compounds,fluorescent compounds, washing solutions, dyes, enzymes, salts, ornucleotides. In some embodiments, a vessel may contain multipledifferent reagents in the vessel (e.g. a buffer, a salt, and an enzymein the same vessel). The combination of multiple reagents in a singlevessel may be a reagent mixture. A reagent mixture may be, for example,in liquid, gel, or lyophilized form. In some embodiments, one or more orall of the vessels in a cartridge are sealed (e.g. a sealed assay unit,reagent unit, etc.). The sealed vessels may be individually sealed, theymay all share the same seal (e.g. a cartridge-wide seal), or groups ofvessels may be sealed together. Sealing materials may be, for example, ametal foil or a synthetic material (e.g. polypropylene). The sealingmaterial may be configured to resist corrosion or degradation. In someembodiments, a vessel may have a septum, such that the contents of thevessel are not exposed to air without puncturing or transversing theseptum.

In some embodiments, a cartridge provided herein may contain all thereagents necessary to perform one or more assays on-board the cartridge.A cartridge may contain all of the reagents on-board necessary toperform 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26,28, 30, 32, 34, 36, 38, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95,100, or more assays. The assays may be any assay or assay type disclosedelsewhere herein. In some embodiments, a cartridge provided herein maycontain within the cartridge all the reagents necessary to perform allof the assays to be performed on a biological sample from a subject. Insome embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22,24, 26, 28, 30, 32, 34, 36, 38, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85,90, 95, 100, or more assays are to be performed in a biological samplefrom a from a subject. A cartridge may also be configured to receive orstore a biological sample from a subject, such that all of the reagentsand biological material necessary to perform one or more assays may beprovided to a device through the insertion of a cartridge containing thesample and reagents into the device. After introduction of a sample intoa device through a cartridge, a sample may be, for example, stored inthe device for archiving or later analysis, or cultured in the device.In some embodiments, all of the reagents in a cartridge are discretelypackaged and/or sealed from interfacing with hardware of a sampleprocessing device.

In some embodiments, provided herein is a system containing a sampleprocessing device and a cartridge. The system, sample processing device,and cartridge may have any of the features described elsewhere herein.The cartridge may be part of the sample processing device. A cartridgemay be positioned in a device or module adjacent to a sensor (e.g. anoptical sensor) or detection station, such that reactions within thecartridge (e.g. in microfluidics channels or vessels in the cartridge)may be measured.

In some embodiments, in systems containing a sample processing deviceand cartridge, the device stores some or all reagents for performingassays within the device. For example, the device may store commonreagents such as water, selected buffers, and detection-relatedcompounds (e.g. chemiluminescent molecules and chromogens) within thedevice. The device may direct reagents for assays to the cartridge asneeded. A device which stores reagents may have tubing to transportreagents from reagent storage locations to the cartridge. Storage ofreagents within the device may, in some situations, increase the speedof reactions or decrease reagent waste.

In other embodiments, in systems containing a sample processing deviceand cartridge, the device does not store any reagents for performingassays within the device. Similarly, in some embodiments, the devicedoes not store any wash solutions or other readily disposable liquids inthe device. In such systems, a cartridge containing all reagentson-board necessary to perform one or more assays may be provided to thedevice. In some embodiments, multiple reagents for performing a singleassay may be provided in a single fluidically isolated vessel (e.g. as areaction mixture). The device may use the reagents provided in thecartridge to perform one or more assays with a biological sample. Thebiological sample may also be included in the cartridge, or it may beseparately provided to the device. In addition, in some embodiments, thedevice may return used reagents to the cartridge, so that all reagentsused for performing one or more assays both enter and leave the devicethrough the cartridge.

A sample processing device which does not store reagents within thedevice (and instead, which receives reagents through the insertion of acartridge or other structure into the device) may have advantages over asample processing device which stores reagents or other disposableswithin or in fluid communication with the device. For example, a sampleprocessing device which stores reagents within the device may requirecomplicated structures for storing and transporting the reagents (e.g.storage areas and tubing). These structures may increase the size of thedevice, require regular maintenance, increase the total amount ofreagents and samples needed to perform assays, and introduce variablesinto assays which may be a source of errors (for example, tubing maylose its shape over time and not deliver accurate volumes). In contrast,a sample processing device which does not store reagents within or influid communication with the device may be smaller, may require lessmaintenance, may use less reagents or sample to perform assays, and mayhave higher accuracy, higher precision, and lower coefficient ofvariation than a device which stores reagents. In another example,typically, devices which store reagents in the device can only contain alimited number of reagents, and thus, can only perform a limited numberof different assays. In addition, such a device may only be configuredto support assays with a limited number of sample types (e.g. only bloodor only urine). Moreover, even if one or more of the reagents in thedevice could be changed to support a different assay, changing of thereagent may be a difficult and time-consuming processing (for example,tubing containing a previous reagent may need to be washed to preventreagent carryover). In contrast, a sample processing device which doesnot store reagents within or in fluid communication with the device maybe capable of performing a higher number of different assays and ofperforming different assays more rapidly, easily, and accurately than adevice which stores reagents, for example due to reduced or eliminatedreagent cross-reactivity or reduced or eliminated human intervention orcalibration).

A bodily fluid collector or any other collection mechanism can bedisposable. For example, a bodily fluid collector can be used once anddisposed. A bodily fluid collector can have one or more disposablecomponents. Alternatively, a bodily fluid collector can be reusable. Thebodily fluid collector can be reused any number of times. In someinstances, the bodily fluid collector can include both reusable anddisposable components. To reduce the environmental impact of disposal,the materials of the cartridge or other bodily fluid collector may bemanufactured of a compostable or other “green” material.

Any component that is inserted into the system or device can beidentified based on identification tags or markings and/or othercommunication means. Based on the identification of such components, thesystem can ensure that said components are suitable for use (e.g., notpassed their expiration date). The system may cross-reference with anon-board and/or remote databases containing data and informationconcerning said components, or a related a protocol or a patient ID.

Components inserted into the system or device may include on-boardssensors. Such sensors may respond to temperature, humidity, light,pressure, vibration, acceleration, and other environmental factors. Suchsensors may be sensitive to absolute levels, durations of exposurelevels, cumulative exposure levels, and other combinations of factors.The system or device can read such sensors and/or communicate with suchsensors when the components are inserted into the system or device orinterface with the user interface to determine how and if the saidcomponent(s) is suitable for use in the system/device based on a set ofrules.

A sample collection unit and/or any other portion of the device may becapable of receiving a single type of sample, or multiple types ofsamples. For example, the sample collection unit may be capable ofreceiving two different types of bodily fluids (e.g., blood, tears). Inanother example, the sample collection unit may be capable of receivingtwo different types of biological samples (e.g., urine sample, stoolsample). Multiple types of samples may or may not be fluids, solids,and/or semi-solids. For example, the sample collection unit may becapable of accepting one or more of, two or more of, or three or more ofa bodily fluid, secretion and/or tissue sample.

A device may be capable of receiving a single type of sample or multipletypes of samples. The device may be capable of processing the singletype of sample or multiple types of samples. In some instances, a singlebodily fluid collector may be used. Alternatively, multiple and/ordifferent bodily fluid collectors may be used.

Sample

A sample may be received by the device. Examples of samples may includevarious fluid samples. In some instances, the sample may be a bodilyfluid sample from the subject. The sample may be an aqueous or gaseoussample. The sample may be a gel. The sample may include one or morefluid component. In some instances, solid or semi-solid samples may beprovided. The sample may include tissue collected from the subject. Thesample may include a bodily fluid, secretion, and/or tissue of asubject. The sample may be a biological sample. The biological samplemay be a bodily fluid, a secretion, and/or a tissue sample. Examples ofbiological samples may include but are not limited to, blood, serum,saliva, urine, gastric and digestive fluid, tears, stool, semen, vaginalfluid, interstitial fluids derived from tumorous tissue, ocular fluids,sweat, mucus, earwax, oil, glandular secretions, breath, spinal fluid,hair, fingernails, skin cells, plasma, nasal swab or nasopharyngealwash, spinal fluid, cerebral spinal fluid, tissue, throat swab, biopsy,placental fluid, amniotic fluid, cord blood, emphatic fluids, cavityfluids, sputum, pus, micropiota, meconium, breast milk and/or otherexcretions. The sample may be provided from a human or animal. Thesample may be provided from a mammal, vertebrate, such as murines,simians, humans, farm animals, sport animals, or pets. The sample may becollected from a living or dead subject.

The sample may be collected fresh from a subject or may have undergonesome form of pre-processing, storage, or transport. The sample may beprovided to a device from a subject without undergoing intervention ormuch time. The subject may contact the device, cartridge, and/or vesselto provide the sample.

A subject may provide a sample, and/or the sample may be collected froma subject. A subject may be a human or animal. The subject may be amammal, vertebrate, such as murines, simians, humans, farm animals,sport animals, or pets. The subject may be living or dead. The subjectmay be a patient, clinical subject, or pre-clinical subject. A subjectmay be undergoing diagnosis, treatment, and/or disease management orlifestyle or preventative care. The subject may or may not be under thecare of a health care professional.

A sample may be collected from the subject by puncturing the skin of thesubject, or without puncturing the skin of the subject. A sample may becollected through an orifice of the subject. A tissue sample may becollected from the subject, whether it be an internal or external tissuesample. The sample may be collected from any portion of the subjectincluding, but not limited to, the subject's finger, hand, arm,shoulder, torso, abdomen, leg, foot, neck, ear, or head.

In some embodiments, the sample may be an environmental sample. Examplesof environmental samples may include air samples, water samples, soilsamples, or plant samples.

Additional samples may include food products, beverages, manufacturingmaterials, textiles, chemicals, therapies, or any other samples.

One type of sample may be accepted and/or processed by the device.Alternatively, multiple types of samples may be accepted and/orprocessed by the device. For example, the device may be capable ofaccepting one or more, two or more, three or more, four or more, five ormore, six or more, seven or more, eight or more, nine or more, ten ormore, twelve or more, fifteen or more, twenty or more, thirty or more,fifty or more, or one hundred or more types of samples. The device maybe capable of accepting and/or processing any of these numbers of sampletypes simultaneously and/or at different times from different or thesame matrices. For example, the device may be capable of preparing,assaying and/or detecting one or multiple types of samples.

Any volume of sample may be provided from the subject or from anothersource. Examples of volumes may include, but are not limited to, about10 mL or less, 5 mL or less, 3 mL or less, 1 μL or less, 500 μL or less,300 μL or less, 250 μL or less, 200 μL or less, 170 μL or less, 150 μLor less, 125 μL or less, 100 μL or less, 75 μL or less, 50 μL or less,25 μL or less, 20 μL or less, 15 μL or less, 10 μL or less, 5 μL orless, 3 μL or less, 1 μL or less, 500 nL or less, 250 nL or less, 100 nLor less, 50 nL or less, 20 nL or less, 10 nL or less, 5 nL or less, 1 nLor less, 500 pL or less, 100 pL or less, 50 pL or less, or 1 pL or less.The amount of sample may be about a drop of a sample. The amount ofsample may be about 1-5 drops of sample, 1-3 drops of sample, 1-2 dropsof sample, or less than a drop of sample. The amount of sample may bethe amount collected from a pricked finger or fingerstick. Any volume,including those described herein, may be provided to the device.

Sample to Device

A sample collection unit may be integral to the device. The samplecollection unit may be separate from the device. In some embodiments,the sample collection unit may be removable and/or insertable from thedevice. The sample collection unit may or may not be provided in acartridge. A cartridge may or may not be removable and/or insertablefrom the device.

A sample collection unit may be configured to receive a sample. Thesample collection unit may be capable of containing and/or confining thesample. The sample collection unit may be capable of conveying thesample to another portion of the device.

The sample collection unit may be in fluid communication with one ormore module of a device. In some instances, the sample collection unitmay be permanent fluid communication with one or more module of thedevice. Alternatively, the sample collection unit may be brought intoand/or out of fluid communication with a module. The sample collectionunit may or may not be selectively fluidically isolated from one or moremodule. In some instances, the sample collection unit may be in fluidcommunication with each of the modules of the device. The samplecollection unit may be in permanent fluid communication with each of themodules, or may be brought into and/or out of fluid communication witheach module.

A sample collection unit may be selectively brought into and/or out offluid communication with one or more modules. The fluid communicationmay be controlled in accordance with one or more protocol or set ofinstructions. A sample collection unit may be brought into fluidcommunication with a first module and out of fluid communication with asecond module, and vice versa.

Similarly, the sample collection unit may be in fluid communication withone or more component of a device. In some instances, the samplecollection unit may be in permanent fluid communication with one or morecomponent of the device. Alternatively, the sample collection unit maybe brought into and/or out of fluid communication with a devicecomponent. The sample collection unit may or may not be selectivelyfluidically isolated from one or more component. In some instances, thesample collection unit may be in fluid communication with each of thecomponents of the device. The sample collection unit may be in permanentfluid communication with each of the components, or may be brought intoand/or out of fluid communication with each component.

One or more mechanisms may be provided for transferring a sample fromthe sample collection unit to a test site. In some embodiments,flow-through mechanisms may be used. For example, a channel or conduitmay connect a sample collection unit with a test site of a module. Thechannel or conduit may or may not have one or more valves or mechanismsthat may selectively permit or obstruct the flow of fluid.

Another mechanism that may be used to transfer a sample from a samplecollection unit to a test site may use one or more fluidically isolatedcomponent. For example, a sample collection unit may provide the sampleto one or more tip or vessel that may be movable within the device. Theone or more tip or vessel may be transferred to one or more module. Insome embodiments, the one or more tip or vessel may be shuttled to oneor more module via a robotic arm or other component of the device. Insome embodiments, the tip or vessel may be received at a module. In someembodiments, a fluid handling mechanism at the module may handle the tipor vessel. For example, a pipette at a module may pick up and/oraspirate a sample provided to the module.

A device may be configured to accept a single sample, or may beconfigured to accept multiple samples. In some instances, the multiplesamples may or may not be multiple types of samples. For example, insome instances a single device may handle a single sample at a time. Forexample, a device may receive a single sample, and may perform one ormore sample processing step, such as a sample preparation step, assaystep, and/or detection step with the sample. The device may completeprocessing or analyzing a sample, before accepting a new sample.

In another example, a device may be capable of handling multiple samplessimultaneously. In one example, the device may receive multiple samplessimultaneously. The multiple samples may or may not be multiple types ofsamples. Alternatively, the device may receive samples in sequence.Samples may be provided to the device one after another, or may beprovided to device after any amount of time has passed. A device may becapable of beginning sample processing on a first sample, receiving asecond sample during said sample processing, and process the secondsample in parallel with the first sample. The first and second samplemay or may not be the same type of sample. The device may be able toparallel process any number of samples, including but not limited tomore than and/or equal to about one sample, two samples, three samples,four samples, five samples, six samples, seven samples, eight samples,nine samples, ten samples, eleven samples, twelve samples, thirteensamples, fourteen samples, fifteen samples, sixteen samples, seventeensamples, eighteen samples, nineteen samples, twenty samples, twenty-fivesamples, thirty samples, forty samples, fifty samples, seventy samples,one hundred samples.

In some embodiments, a device may comprise one, two or more modules thatmay be capable of processing one, two or more samples in parallel. Thenumber of samples that can be processed in parallel may be determined bythe number of available modules and/or components in the device.

When a plurality of samples is being processed simultaneously, thesamples may begin and/or end processing at any time. The samples neednot begin and/or end processing at the same time. A first sample mayhave completed processing while a second sample is still beingprocessed. The second sample may begin processing after the first samplehas begun processing. As samples have completed processing, additionalsamples may be added to the device. In some instances, the device may becapable of running continuously with samples being added to the deviceas various samples have completed processing.

The multiple samples may be provided simultaneously. The multiplesamples may or may not be the same type of sample. For example, multiplesample collection units may be provided to a device. For example, one,two or more lancets may be provided on a device or may be brought intofluid communication with a sample collection unit of a device. Themultiple sample collection units may receive samples simultaneously orat different times. Multiple of any of the sample collection mechanismsdescribed herein may be used. The same type of sample collectionmechanisms, or different types of sample collection mechanisms may beused.

The multiple samples may be provided in sequence. In some instances,multiple sample collection units, or single sample collection units maybe used. Any combination of sample collection mechanisms describedherein may be used. A device may accept one sample at a time, twosamples at a time, or more. Samples may be provided to the device afterany amount of time has elapsed.

Modules

Devices may comprise one or more module. A module may be capable ofperforming one or more, two or more, or all three of a samplepreparation step, assay step, and/or detection step. FIG. 3 shows anexample of a module 300. A module may comprise one or more, two or more,or three or more of a sample preparation station 310, and/or an assaystation 320, and/or a detection station 330. In some embodiments,multiple of a sample preparation station, assay station, and/ordetection station are provided. A module may also include a fluidhandling system 340.

A module may include one or more sample preparation station. A samplepreparation station may include one or more component configured forchemical processing and/or physical processing. Examples of such samplepreparation processes may include dilution, concentration/enrichment,separation, sorting, filtering, lysing, chromatography, incubating, orany other sample preparation step. A sample preparation station mayinclude one or more sample preparation components, such as a separationsystem (including, but not limited to, a centrifuge), magnets (or othermagnetic field-inducing devices) for magnetic separation, a filter, aheater, or diluents.

A sample preparation station may be insertable into or removable from asystem, device, or module. A sample preparation station may comprise acartridge. In some embodiments, any description of a cartridge providedherein may apply to a sample preparation station, and vice-versa.

One or more assay station may be provided to a module. The assay stationmay include one or more component configured to perform one or more ofthe following assays or steps: immunoassay, nucleic acid assay,receptor-based assay, cytometric assay, colorimetric assay, enzymaticassay, electrophoretic assay, electrochemical assay, spectroscopicassay, chromatographic assay, microscopic assay, topographic assay,calorimetric assay, turbidimetric assay, agglutination assay,radioisotope assay, viscometric assay, coagulation assay, clotting timeassay, protein synthesis assay, histological assay, culture assay,osmolarity assay, and/or other types of assays or combinations thereof.The assay station may be configured for proteinaceous assay, includingimmunoassay and Enzymatic assay or any other assay that involvesinteraction with a proteinaeous component. Topographic assays in somecases include morphological assays. Examples of other components thatmay be included in an assay station or a module are, without limitation,one or more of the following: temperature control unit, heater, thermalblock, cytometer, electromagnetic energy source (e.g., x-ray, lightsource), assay units, reagent units, and/or supports. In someembodiments, a module includes one or more assay stations capable ofperforming nucleic acid assay and proteinaceous assay (includingimmunoassay and enzymatic assay). In some embodiments, a module includesone or more assay stations capable of performing fluorescent assay andcytometry.

An assay station may be insertable into or removable from a system,device, or module. An assay station may comprise a cartridge. In someembodiments, any description of an assay/reagent unit support orcartridge provided herein may apply to an assay station, and vice-versa.

In some embodiments, a system, device, or module provided herein mayhave an assay station/cartridge receiving location. The assay stationreceiving location may be configured to receive a removable orinsertable assay station. The assay station receiving location may besituated in the module, device, or system such that an assay stationpositioned in the receiving location (and assay units therein) may beaccessible by a sample handling system of the module, device, or system.The assay station receiving location may be configured to position anassay station at a precise location within the receiving location, suchthat a sample handling system may accurately access components of theassay station. An assay station receiving location may be a tray. Thetray may be movable, and may have multiple positions, for example, afirst position where the tray extends outside of the housing of thedevice, and a second position wherein the tray is inside of the housingof the device. In some embodiments, an assay station may be locked inplace in an assay station receiving location. In some embodiments, theassay station receiving location may contain or be operatively coupledto a thermal control unit to regulate the temperature of the assaystation. In some embodiments, the assay station receiving location maycontain or be operatively coupled to a detector (e.g. bar code detector,RFID detector) for an identifier (e.g. bar code, RFID tag) which may beon an assay station. The identifier detector may be in communicationwith a controller or other component of the device, such that theidentifier detector can transmit information regarding the identity ofan assay station/cartridge inserted into the device to the device orsystem controller.

The assay station may or may not be located separately from thepreparation station. In some instances, an assay station may beintegrated within the preparation station. Alternatively, they may bedistinct stations, and a sample or other substance may be transmittedfrom one station to another.

Assay units may be provided, and may have one or more characteristics asdescribed further elsewhere herein. Assay units may be capable ofaccepting and/or confining a sample. The assay units may be fluidicallyisolated from or hydraulically independent of one another. In someembodiments, assay units may have a tip format. An assay tip may have aninterior surface and an exterior surface. The assay tip may have a firstopen end and a second open end. In some embodiments, assay units may beprovided as an array. Assay units may be movable. In some embodiments,individual assay units may be movable relative to one another and/orother components of the device. In some instances, one or a plurality ofassay units may be moved simultaneously. In some embodiments, an assayunit may have a reagent or other reactant coated on a surface. In someembodiments, a succession of reagents may be coated or deposited on asurface, such as a tip surface, and the succession of reagents can beused for sequential reactions. Alternatively, assay units may containbeads or other surfaces with reagents or other reactants coated thereonor absorbed, adsorbed or adhered therein. In another example, assayunits may contain beads or other surfaces coated with or formed ofreagents or other reactants that may dissolve. In some embodiments,assay units may be cuvettes. In some instances, cuvettes may beconfigured for cytometry, may include microscopy cuvettes.

Reagent units may be provided and may have one or more characteristicsas described further elsewhere herein. Reagent units may be capable ofaccepting and/or confining a reagent or a sample. Reagent units may befluidically isolated from or hydraulically independent of one another.In some embodiments, reagent units may have a vessel format. A reagentvessel may have an interior surface and an exterior surface. The reagentunit may have an open end and a closed end. In some embodiments, thereagent units may be provided as an array. Reagent units may be movable.In some embodiments, individual reagent units may be movable relative toone another and/or other components of the device. In some instances,one or a plurality of reagent units may be moved simultaneously. Areagent unit can be configured to accept one or more assay unit. Thereagent unit may have an interior region into which an assay unit can beat least partially inserted.

A support may be provided for the assay units and/or reagent units. Insome embodiments, the support may have an assay station format, acartridge format or a microcard format. In some embodiments a supportmay have a patch format or may be integrated into a patch or animplantable sensing an analytical unit. One or more assay/reagent unitsupport may be provided within a module. The support may be shaped tohold one or more assay units and/or reagent units. The support may keepthe assay units and/or reagent units aligned in a vertical orientation.The support may permit assay units and/or reagent units to be moved ormovable. Assay units and/or reagent units may be removed from and/orplaced on a support. The device and/or system may incorporate one ormore characteristics, components, features, or steps provided in U.S.Patent Publication No. 2009/0088336, which is hereby incorporated byreference in its entirety.

A module may include one or more detection stations. A detection stationmay include one or more sensors that may detect visual/optical signals,infra-red signals, heat/temperature signals, ultraviolet signals, anysignal along an electromagnetic spectra, electric signals, chemicalsignals, audio signals, pressure signals, motion signals, or any othertype of detectable signals. The sensors provided herein may or may notinclude any of the other sensors described elsewhere herein. Thedetection station may be located separately from the sample preparationand/or assay station. Alternatively, the detection station may belocated in an integrated manner with the sample preparation and/or assaystation. A detection station may contain one or more detection units,including any detection unit disclosed elsewhere herein. A detectionstation may contain, for example, a spectrophotometer, a PMT, aphotodiode, a camera, an imaging device, a CCD or CMOS optical sensor,or a non-optical sensor. In some embodiments, a detection station maycontain a light source and optical sensor. In some embodiments, adetection station may contain a microscope objective and an imagingdevice.

In some embodiments, a sample may be provided to one or more samplepreparation station before being provided to an assay station. In someinstances, a sample may be provided to a sample preparation after beingprovided to an assay station. A sample may undergo detection before,during, or after it is provided to a sample preparation station and/orassay station.

A fluid handling system may be provided to a module. The fluid handlingsystem may permit the movement of a sample, reagent, or a fluid. Thefluid handling system may permit the dispensing and/or aspiration of afluid. The fluid handling system may pick up a desired fluid from aselected location and/or may dispense a fluid at a selected location.The fluid handling system may permit the mixing and/or reaction of twoor more fluids. In some cases, a fluid handling mechanism may be apipette. Examples of pipettes or fluid handling mechanisms are providedin greater detail elsewhere herein.

Any description herein of a fluid handling system may also apply toother sample handling systems, and vice versa. For example, a samplehandling system may transport any type of sample, including but notlimited to bodily fluids, secretions, or tissue samples. A samplehandling system may be capable of handling fluids, solids, orsemi-solids. A sample handling system may be capable of accepting,depositing, and/or moving a sample, and/or any other substance withinthe device may be useful and/or necessary for sample processing withinthe device. A sample handling system may be capable of accepting,depositing, and/or moving a container (e.g., assay unit, reagent unit)that may contain a sample, and/or any other substance within the device.

A fluid handling system may include a tip. For example, a pipette tipmay be removably connected to a pipette. The tip may interface with apipette nozzle. Examples of tip/nozzle interfaces are provided ingreater detail elsewhere herein.

Another example of a fluid handling system may use flow-through designs.For example, a fluid handling system may incorporate one or morechannels and/or conduits through which a fluid may flow. The channel orconduit may comprise one or more valves that may selectively stop and/orpermit the flow of fluid.

A fluid handling system may have one or more portion that may result influid isolation. For example, a fluid handling system may use a pipettetip that may be fluidically isolated from other components of thedevice. The fluidically isolated portions may be movable. In someembodiments, the fluid handling system tips may be assay tips asdescribed elsewhere herein.

A module may have a housing and/or support structure. In someembodiments, a module may have a support structure upon which one ormore component of the module may rest. The support structure may supportthe weight of one or more component of the module. The components may beprovided above the support structure, on the side of the supportstructure, and/or under the support structure. The support structure maybe a substrate which may connect and/or support various components ofthe module. The support structure may support one or more samplepreparation station, assay station, and/or detection station of themodule. A module may be self-contained. The modules may be movedtogether. The various components of the module may be capable of beingmoved together. The various components of the module may be connected toone another. The components of the module may share a common support.

A module may be enclosed or open. A housing of the module may enclosethe module therein. The housing may completely enclose the module or maypartially enclose the module. The housing may form an air-tightenclosure around the module. Alternatively, the housing need not beair-tight. The housing may enable the temperature, humidity, pressure,or other characteristics within the module or component(s) of the moduleto be controlled.

Electrical connections may be provided for a module. A module may beelectrically connected to the rest of the device. A plurality of modulesmay or may not be electrically connected to one another. A module may bebrought into electrical connection with a device when a module isinserted/attached to the device. The device may provide power (orelectricity) to the module. A module may be disconnected from theelectrical source when removed from the device. In one instance, when amodule is inserted into the device, the module makes an electricalconnection with the rest of the device. For example, the module may pluginto the support of a device. In some instances, the support (e.g.,housing) of the device may provide electricity and/or power to themodule.

A module may also be capable of forming fluidic connections with therest of the device. In one example, a module may be fluidicallyconnected to the rest of the device. Alternatively, the module may bebrought into fluidic communication with the rest of the device via,e.g., a fluid handling system disclosed herein. The module may bebrought into fluidic communication when the module is inserted/attachedto the device, or may be selectively brought into fluidic communicationanytime after the module is inserted/attached to the device. A modulemay be disconnected from fluidic communication with the device when themodule is removed from the device and/or selectively while the module isattached to the device. In one example, a module may be in or may bebrought into fluidic communication with a sample collection unit of thedevice. In another example, a module may be in or may be brought intofluidic communication with other modules of the device.

A module may have any size or shape, including those described elsewhereherein. A module may have a size that is equal to, or smaller than thedevice. The device module may enclose a total volume of less than orequal to about 4 m³, 3 m³, 2.5 m³, 2 m³, 1.5 m³, 1 m³, 0.75 m³, 0.5 m³,0.3 m³, 0.2 m³, 0.1 m³, 0.08 m³, 0.05 m³, 0.03 m³, 0.01 m³, 0.005 m³,0.001 m³, 500 cm³, 100 cm³, 50 cm³, 10 cm³, 5 cm³, 1 cm³, 0.5 cm³, 0.1cm³, 0.05 cm³, 0.01 cm³, 0.005 cm³, or 0.001 cm³. The module may haveany of the volumes described elsewhere herein.

The module and/or module housing may have a footprint covering a lateralarea of the device. In some embodiments, the device footprint may beless than or equal to about 4 m², 3 m², 2.5 m², 2 m², 1.5 m², 1 m², 0.75m², 0.5 m², 0.3 m², 0.2 m², 0.1 m², 0.08 m², 0.05 m², 0.03 m², 100 cm²,80 cm², 70 cm², 60 cm², 50 cm², 40 cm², 30 cm², 20 cm², 15 cm², 10 cm²,7 cm², 5 cm², 1 cm², 0.5 cm², 0.1 cm², 0.05 cm², 0.01 cm, 0.005 cm², or0.001 cm².

The module and/or module housing may have a lateral dimension (e.g.,width, length, or diameter) or a height less than or equal to about 4 m,3 m, 2.5 m, 2 m, 1.5 m, 1.2 m, 1 m, 80 cm, 70 cm, 60 cm, 50 cm, 40 cm,30 cm, 25 cm, 20 cm, 15 cm, 12 cm, 10 cm, 8 cm, 5 cm, 3 cm, 1 cm, 0.5cm, 0.1 cm, 0.05 cm, 0.01 cm, 0.005 cm, or 0.001 cm. The lateraldimensions and/or height may vary from one another. Alternatively, theymay be the same. In some instances, the module may be tall and thin, ormay be short and squat. The height to lateral dimension ratio may begreater than or equal to 100:1, 50:1, 30:1, 20:1, 10:1, 9:1, 8:1, 7:1,6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9,1:10, 1:20, 1:30, 1:50, or 1:100. The module and/or the module housingmay proportionally be tall and thin.

The module and/or module housing may have any shape. In someembodiments, the module may have a lateral cross-sectional shape of arectangle or square. In other embodiments, the module may have a lateralcross-sectional shape of a circle, ellipse, triangle, trapezoid,parallelogram, pentagon, hexagon, octagon, or any other shape. Themodule may have a vertical cross-sectional shape of a circle, ellipse,triangle, rectangle, square, trapezoid, parallelogram, pentagon,hexagon, octagon, or any other shape. The module may or may not have abox-like shape.

Any number of modules may be provided for a device. A device may beconfigured to accept a fixed number of modules. Alternatively, thedevice may be configured to accept a variable number of modules. In someembodiments, each module for the device may have the same componentsand/or configurations. Alternatively, different modules for the devicemay have varying components and/or configurations. In some instances,the different modules may have the same housing and/or support structureformats. In another example, the different modules may still have thesame overall dimensions. Alternatively, they may have varyingdimensions.

In some instances a device may have a single module. The single modulemay be configured to accept a single sample at once, or may be capableof accepting a plurality of samples simultaneously or in sequence. Thesingle module may be capable of performing one or more samplepreparation step, assay step, and/or detection step. The single modulemay or may not be swapped out to provide different functionality.

Further details and descriptions of modules and module components aredescribed further elsewhere herein. Any such embodiments of such modulesmay be provided in combination with others or alone.

Racks

In an aspect of the invention, a system having a plurality of modules isprovided. The system is configured to assay a biological sample, such asa fluid and/or tissue sample from a subject.

In some embodiments, the system comprises a plurality of modules mountedon a support structure. In an embodiment, the support structure is arack having a plurality of mounting stations, an individual mountingstation of the plurality of mounting stations for supporting a module.

In an embodiment, the rack comprises a controller communicativelycoupled to the plurality of modules. In some situations, the controlleris communicatively coupled to a fluid handling system, as describedbelow. The controller is configured to control the operation of themodules to prepare and/or process a sample, such as to assay a samplevia one or more of the techniques described herein.

An individual module of the plurality of modules comprises a samplepreparation station, assay station, and/or detection station. The systemis configured to perform (a) multiple sample preparation proceduresselected from the group consisting of sample processing, centrifugation,separation, physical separation and chemical separation, and (b) atleast one type of assay selected from the group consisting ofimmunoassay, nucleic acid assay, receptor-based assay, cytometric assay,colorimetric assay, enzymatic assay, electrophoretic assay,electrochemical assay, spectroscopic assay, chromatographic assay,microscopic assay, topographic assay, calorimetric assay, turbidimetricassay, agglutination assay, radioisotope assay, viscometric assay,coagulation assay, clotting time assay, protein synthesis assay,histological assay, culture assay, osmolarity assay, and/or other typesof assays or combinations thereof. In some embodiments, separationincludes magnetic separation, such as, e.g., separation with the aid ofa magnetic field.

In an embodiment, the support structure is a rack-type structure forremovably holding or securing an individual module of the plurality ofmodules. The rack-type structure includes a plurality of bays configuredto accept and removably secure a module. In one example, as shown inFIG. 4, a rack 400 may have one or more modules 410 a, 410 b, 410 c, 410d, 410 e, 410 f. The modules may have a vertical arrangement where theyare positioned over one another. For example, six modules may be stackedon top of one another. The modules may have a horizontal arrangementwhere they are adjacent to one another. In another example, the modulesmay form an array. FIG. 5 illustrates an example of a rack 500 having aplurality of modules 510 that form an array. For example, the modulesmay form a vertical array that is M modules high and/or N modules wide,wherein M, N are positive whole numbers. In other embodiments, a rackmay support an array of modules, where a horizontal array of modules isformed. For example, the modules may form a horizontal array that is Nmodules wide and/or P modules long, wherein N and P are positive wholenumbers. In another example, a three-dimensional array of modules may besupported by a rack, where the modules form a block that is M moduleshigh, N modules wide, and P modules long, where M, N, and P are positivewhole numbers. A rack may be able to support any number of moduleshaving any number of configurations.

In some embodiments, racks may have one or more bays, each bayconfigured to accept one or more module. A device may be capable ofoperating when a bay has accepted a module. A device may be capable ofoperating even if one or more bays have not accepted a module.

FIG. 6 shows another embodiment of a rack mounting configuration. One ormore module 600 a, 600 b may be provided adjacent to one another. Anynumbers of modules may be provided. For example, the modules may bevertically stacked atop one another. For instances, N modules may bevertically stacked on top of one another, where N is any positive wholenumber. In another example, the modules may be horizontally connected toone another. Any combination of vertical and/or horizontal connectionsbetween modules may be provided. The modules may directly contact oneanother or may have a connecting interface. In some instances, modulesmay be added or removed from the stack/group. The configuration may becapable of accommodating any number of modules. In some embodiments, thenumber of modules may or may not be restricted by a device housing.

In another embodiment, the support structure is disposed below a firstmodule and successive modules are mountable on one another with orwithout the aid of mounting members disposed on each module. Themounting members may be connecting interfaces between modules. In anexample, each module includes a magnetic mounting structure for securinga top surface of a first module to a bottom surface to a second module.Other connecting interfaces may be employed, which may include magneticfeatures, adhesives, sliding features, locking features, ties,snap-fits, hook-and-loop fasteners, twisting features, or plugs. Themodules may be mechanically and/or electrically connected to oneanother. In such fashion, modules may be stacked on one or next toanother to form a system for assaying a sample.

In other embodiments, a system for assaying a sample comprises a housingand a plurality of modules within the housing. In an embodiment, thehousing is a rack having a plurality of mounting stations, an individualmounting station of the plurality of mounting stations for supporting amodule. For example, a rack may be integrally formed with the housing.Alternatively, the housing may contain or surround the rack. The housingand the rack may or may not be formed of separate pieces that may or maynot be connected to one another. An individual module of the pluralityof modules comprises at least one station selected from the groupconsisting of a sample preparation station, assay station and detectionstation. The system comprises a fluid handling system configured totransfer a sample or reagent vessel within the individual module or fromthe individual module to another module within the housing of thesystem. In an embodiment, the fluid handling system is a pipette.

In some embodiments, all modules could be shared within a device orbetween devices. For example, a device may have one, some or all of itsmodules as specialized modules. In this case, a sample may betransported from one module to another module as need be. This movementmay be sequential or random.

Any of the modules can be a shared resource or may comprise designatedshared resources. In one example a designated shared resource may be aresource not available to all modules, or that may be available inlimited numbers of modules. A shared resource may or may not beremovable from the device. An example of a shared resource may include acytometry station.

In an embodiment, the system further comprises a cytometry station forperforming cytometry on one or more samples. The cytometry station maybe supported by the rack and operatively coupled to each of theplurality of modules by a sample handling system.

Cytometry assays are typically used to optically measure characteristicsof individual cells. The cells being monitored may be live and/or deadcells. By using appropriate dyes, stains, or other labeling molecules,cytometry may be used to determine the presence, quantity, and/ormodifications of specific proteins, nucleic acids, lipids,carbohydrates, or other molecules. Properties that may be measured bycytometry also include measures of cellular function or activity,including but not limited to phagocytosis, active transport of smallmolecules, mitosis or meiosis; protein translation, gene transcription,DNA replication, DNA repair, protein secretion, apoptosis, chemotaxis,mobility, adhesion, antioxidizing activity, RNAi, protein or nucleicacid degradation, drug responses, infectiousness, and the activity ofspecific pathways or enzymes. Cytometry may also be used to determineinformation about a population of cells, including but not limited tocell counts, percent of total population, and variation in the samplepopulation for any of the characteristics described above. The assaysdescribed herein may be used to measure one or more of the abovecharacteristics for each cell, which may be advantageous to determiningcorrelations or other relationships between different characteristics.The assays described herein may also be used to independently measuremultiple populations of cells, for example by labeling a mixed cellpopulation with antibodies specific for different cell lines.

Cytometry may be useful for determining characteristics of cells inreal-time. Characteristics of cells may be monitored continuously and/orat different points in time. The different points in time may be atregular or irregular time intervals. The different points in time may bein accordance with a predetermined schedule or may be triggered by oneor more event. Cytometry may use one or more imaging or other sensingtechnique described herein to detect change in cells over time. This mayinclude cell movement or morphology. Kinematics or dynamics of a samplemay be analyzed. Time series analysis may be provided for the cells.Such real-time detection may be useful for calculation of agglutination,coagulation, or prothrombin time. The presence of one or more moleculeand/or disease, response to a disease and/or drug, may be ascertainedbased on the time-based analysis.

In an example, cytometric analysis is by flow cytometry or bymicroscopy. Flow cytometry typically uses a mobile liquid medium thatsequentially carries individual cells to an optical detector. Microscopytypically uses optical means to detect stationary cells, generally byrecording at least one magnified image. For microscopy, the stationarycells may be in a microscopy cuvette or slide, which may be positionedon a microscopy stage adjacent to or in optical connection with animaging device for detecting the cells. Imaged cells may be, forexample, counted or measured for one or more antigens or other features.It should be understood that flow cytometry and microscopy are notentirely exclusive. As an example, flow cytometry assays use microscopyto record images of cells passing by the optical detector. Many of thetargets, reagents, assays, and detection methods may be the same forflow cytometry and microscopy. As such, unless otherwise specified, thedescriptions provided herein should be taken to apply to these and otherforms of cytometric analyses known in the art.

In some embodiments, up to about 10,000 cells of any given type may bemeasured. In other embodiments, various numbers of cells of any giventype are measured, including, but not limited to, more than, and/orequal to about 10 cells, 30 cells, 50 cells, 100 cells, 150 cells, 200cells, 300 cells, 500 cells, 700 cells, 1000 cells, 1500 cells, 2000cells, 3000 cells, 5000 cells, 6000 cells, 7000 cells, 8000 cells, 9000cells, 10000 cells, 100,000 cells, 500,000 cells, 1,000,000 cells,5,000,000 cells, or 10,000,000 cells.

In some embodiments, cytometry is performed in microfluidic channels.For instance, flow cytometry analyses are performed in a single channelor in parallel in multiple channels. In some embodiments, flow cytometrysequentially or simultaneously measures multiple cell characteristics.In some instances, cytometry may occur within one or more of thetips/vessels described herein. Cytometry may be combined with cellsorting, where detection of cells that fulfill a specific set ofcharacteristics are diverted from the flow stream and collected forstorage, additional analysis, and/or processing. Such sorting mayseparate multiple populations of cells based on different sets ofcharacteristics, such as 3 or 4-way sorting.

FIG. 7 shows a system 700 having a plurality of modules 701-706 and acytometry station 707, in accordance with an embodiment of theinvention. The plurality of modules include a first module 701, secondmodule 702, third module 703, fourth module 704, fifth module 705 andsixth module 706.

The cytometry station 707 is operatively coupled to each of theplurality of modules 701-706 by way of a sample handling system 708. Thesample handling system 708 may include a pipette, such as a positivedisplacement, air displacement or suction-type pipette, as describedherein.

The cytometry station 707 includes a cytometer for performing cytometryon a sample, as described above and in other embodiments of theinvention. The cytometry station 707 may perform cytometry on a samplewhile one or more of the modules 701-706 perform other preparationand/or assaying procedure on another sample. In some situations, thecytometry station 707 performs cytometry on a sample after the samplehas undergone sample preparation in one or more of the modules 701-706.

The system 700 includes a support structure 709 having a plurality ofbays (or mounting stations). The plurality of bays is for docking themodules 701-706 to the support structure 709. The support structure 709,as illustrated, is a rack.

Each module is secured to rack 709 with the aid of an attachment member.In an embodiment, an attachment member is a hook fastened to either themodule or the bay. In such a case, the hook is configured to slide intoa receptacle of either the module or the bay. In another embodiment, anattachment member includes a fastener, such as a screw fastener. Inanother embodiment, an attachment member is formed of a magneticmaterial. In such a case, the module and bay may include magneticmaterials of opposite polarities so as to provide an attractive force tosecure the module to the bay. In another embodiment, the attachmentmember includes one or more tracks or rails in the bay. In such a case,a module includes one or more structures for mating with the one or moretracks or rails, thereby securing the module to the rack 709.Optionally, power may be provided by the rails.

An example of a structure that may permit a module to mate with a rackmay include one or more pins. In some cases, modules receive powerdirectly from the rack. In some cases, a module may be a power sourcelike a lithion ion, or fuel cell powered battery that powers the deviceinternally. In an example, the modules are configured to mate with therack with the aid of rails, and power for the modules comes directlyfrom the rails. In another example, the modules mate with the rack withthe aid of attachment members (rails, pins, hooks, fasteners), but poweris provided to the modules wirelessly, such as inductively (i.e.,inductive coupling).

In some embodiments, a module mating with a rack need not require pins.For example, an inductive electrical communication may be providedbetween the module and rack or other support. In some instances,wireless communications may be used, such as with the aid of ZigBeecommunications or other communication protocols.

Each module may be removable from the rack 709. In some situations, onemodule is replaceable with a like, similar or different module. In anembodiment, a module is removed from the rack 709 by sliding the moduleout of the rack. In another embodiment, a module is removed from therack 709 by twisting or turning the module such that an attachmentmember of the module disengages from the rack 709. Removing a modulefrom the rack 709 may terminate any electrical connectivity between themodule and the rack 709.

In an embodiment, a module is attached to the rack by sliding the moduleinto the bay. In another embodiment, a module is attached to the rack bytwisting or turning the module such that an attachment member of themodule engages the rack 709. Attaching a module to the rack 709 mayestablish an electrical connection between the module and the rack. Theelectrical connection may be for providing power to the module or to therack or to the device from the module and/or providing a communicationsbus between the module and one or more other modules or a controller ofthe system 700.

Each bay of the rack may be occupied or unoccupied. As illustrated, allbays of the rack 709 are occupied with a module. In some situations,however, one or more of the bays of the rack 709 are not occupied by amodule. In an example, the first module 701 has been removed from therack. The system 700 in such a case may operate without the removedmodule.

In some situations, a bay may be configured to accept a subset of thetypes of modules the system 700 is configured to use. For example, a baymay be configured to accept a module capable of running an agglutinationassay but not a cytometry assay. In such a case, the module may be“specialized” for agglutination. Agglutination may be measured in avariety of ways. Measuring the time-dependent change in turbidity of thesample is one method. One can achieve this by illuminating the samplewith light and measuring the reflected light at 90 degrees with anoptical sensor, such as a photodiode or camera. Over time, the measuredlight would increase as more light is scattered by the sample. Measuringthe time dependent change in transmittance is another example. In thelatter case, this can be achieved by illuminating the sample in a vesseland measuring the light that passes through the sample with an opticalsensor, such as a photodiode or a camera. Over time, as the sampleagglutinates, the measured light may reduce or increase (depending, forexample, on whether the agglutinated material remains in suspension orsettles out of suspension). In other situations, a bay may be configuredto accept all types of modules that the system 700 is configured to use,ranging from detection stations to the supporting electrical systems.

Each of the modules may be configured to function (or perform)independently from the other modules. In an example, the first module701 is configured to perform independently from the second 702, third703, fourth 704, fifth 705 and sixth 706 modules. In other situations, amodule is configured to perform with one or more other modules. In sucha case, the modules may enable parallel processing of one or moresamples. In an example, while the first module 701 prepares a sample,the second module 702 assays the same or different sample. This mayenable a minimization or elimination of downtime among the modules.

The support structure (or rack) 709 may have a server typeconfiguration. In some situations, various dimensions of the rack arestandardized. In an example, spacing between the modules 701-706 isstandardized as multiples of at least about 0.5 inches, or 1 inch, or 2inches, or 3 inches, or 4 inches, or 5 inches, or 6 inches, or 7 inches,or 8 inches, or 9 inches, or 10 inches, or 11 inches, or 12 inches.

The rack 709 may support the weight of one or more of the modules701-706. Additionally, the rack 709 has a center of gravity that isselected such that the module 701 (top) is mounted on the rack 709without generating a moment arm that may cause the rack 709 to spin orfall over. In some situations, the center of gravity of the rack 709 isdisposed between the vertical midpoint of the rack and a base of therack, the vertical midpoint being 50% from the base of the rack 709 anda top of the rack. In an embodiment, the center of gravity of the rack709, as measured along a vertical axis away from the base of the rack709, is disposed at least about 0.1%, or 1%, or 10%, or 20%, or 30%, or40%, or 50%, or 60%, or 70%, or 80%, or 90%, or 100% of the height ofthe rack as measured from the base of the rack 709.

A rack may have multiple bays (or mounting stations) configured toaccept one or more modules. In an example, the rack 709 has six mountingstations for permitting each of the modules 701-706 to mount the rack.In some situations, the bays are on the same side of the rack. In othersituations, the bays are on alternating sides of the rack.

In some embodiments, the system 700 includes an electrical connectivitycomponent for electrically connecting the modules 701-706 to oneanother. The electrical connectivity component may be a bus, such as asystem bus. In some situations, the electrical connectivity componentalso enables the modules 701-706 to communicate with each other and/or acontroller of the system 700.

In some embodiments, the system 700 includes a controller (not shown)for facilitating processing of samples with the aid of one or more ofthe modules 701-706. In an embodiment, the controller facilitatesparallel processing of the samples in the modules 701-706. In anexample, the controller directs the sample handling system 708 toprovide a sample in the first module 701 and second module 702 to rundifferent assays on the sample at the same time. In another example, thecontroller directs the sample handling system 708 to provide a sample inone of the modules 701-706 and also provide the sample (such as aportion of a finite volume of the sample) to the cytometry station 707so that cytometry and one or more other sample preparation proceduresand/or assays are done on the sample in parallel. In such fashion, thesystem minimizes, if not eliminates, downtime among the modules 701-706and the cytometry station 707.

Each individual module of the plurality of modules may include a samplehandling system for providing samples to and removing samples fromvarious processing and assaying modules of the individual module. Inaddition, each module may include various sample processing and/orassaying modules, in addition to other components for facilitatingprocessing and/or assaying of a sample with the aid of the module. Thesample handling system of each module may be separate from the samplehandling system 708 of the system 700. That is, the sample handlingsystem 708 transfers samples to and from the modules 701-706, whereasthe sample handling system of each module transfers samples to and fromvarious sample processing and/or assaying modules included within eachmodule.

In the illustrated example of FIG. 7, the sixth module 706 includes asample handling system 710 including a suction-type pipette 711 andpositive displacement pipette 712. The sixth module 706 includes acentrifuge 713, a spectrophotometer 714, a nucleic acid assay (such as apolymerase chain reaction (PCR) assay) station 715 and PMT 716. Anexample of the spectrophotometer 714 is shown in FIG. 70 (see below).The sixth module 706 further includes a cartridge 717 for holding aplurality of tips for facilitating sample transfer to and from eachprocessing or assaying module of the sixth module.

In an embodiment, the suction type pipette 711 includes 1 or more, or 2or more, or 3 or more, or 4 or more, or 5 or more, or 6 or more, or 7 ormore, or 8 or more, or 9 or more, or 10 or more, or 15 or more, or 20 ormore, or 30 or more, or 40 or more, or 50 or more heads. In an example,the suction type pipette 711 is an 8-head pipette with eight heads. Thesuction type pipette 711 may be as described in other embodiments of theinvention.

In some embodiments, the positive displacement pipette 712 has acoefficient of variation less than or equal to about 20%, 15%, 12%, 10%,9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.3%, or 0.1% or less. Thecoefficient of variation is determined according to σ/μ wherein ‘σ’ isthe standard deviation and ‘μ’ is the mean across sample measurements.

In an embodiment, all modules are identical to one another. In anotherembodiment, at least some of the modules are different from one another.In an example, the first, second, third, fourth, fifth, and sixthmodules 701-706 include a positive displacement pipette and suction-typepipette and various assays, such as a nucleic acid assay andspectrophotometer. In another example, at least one of the modules701-706 may have assays and/or sample preparation stations that aredifferent from the other modules. In an example, the first module 701includes an agglutination assay but not a nucleic acid amplificationassay, and the second module 702 includes a nucleic acid assay but notan agglutination assay. Modules may not include any assays.

In the illustrated example of FIG. 7, the modules 701-706 include thesame assays and sample preparation (or manipulation) stations. However,in other embodiments, each module includes any number and combination ofassays and processing stations described herein.

The modules may be stacked vertically or horizontally with respect toone another. Two modules are oriented vertically in relation to oneanother if they are oriented along a plane that is parallel,substantially parallel, or nearly parallel to the gravitationalacceleration vector. Two modules are oriented horizontally in relationto one another if they are oriented along a plane orthogonal,substantially orthogonal, or nearly orthogonal to the gravitationalacceleration vector.

In an embodiment, the modules are stacked vertically, i.e., one moduleon top of another module. In the illustrated example of FIG. 7, the rack709 is oriented such that the modules 701-706 are disposed vertically inrelation to one another. However, in other situations the modules aredisposed horizontally in relation to one another. In such a case, therack 709 may be oriented such that the modules 701-706 may be situatedhorizontally alongside one another.

Referring now to FIG. 7A, yet another embodiment of a system 730 isshown with a plurality of modules 701 to 704. This embodiment of FIG. 7Ashows a horizontal configuration wherein the modules 701 to 704 aremounted to a support structure 732 on which a transport device 734 canmove along the X, Y, and/or optionally Z axis to move elements such asbut not limited sample vessels, tips, cuvettes, or the like within amodule and/or between modules. By way of non-limiting example, themodules 701-704 are oriented horizontally in relation to one another ifthey are oriented along a plane orthogonal, substantially orthogonal, ornearly orthogonal to the gravitational acceleration vector.

It should be understood that, like the embodiment of FIG. 7, modules701-704 may all be modules that are identical to one another. In anotherembodiment, at least some of the modules are different from one another.In an example, the first, second, third, and/or fourth modules 701-704may be replaced by one or more other modules that can occupy thelocation of the module being replaced. The other modules may optionallyprovide different functionality such as but not limited to a replacingone of the modules 701-704 with one or more cytometry modules 707,communications modules, storage modules, sample preparation modules,slide preparation modules, tissue preparation modules, or the like. Forexample, one of the modules 701-704 may be replaced with one or moremodules that provide a different hardware configuration such as but notlimited to provide a thermal controlled storage chamber for incubation,storage between testing, and/or storage after testing. Optionally, themodule replacing one or more of the modules 701-704 can provide anon-assay related functionality, such as but not limited to additionaltelecommunication equipment for the system 730, additional imaging oruser interface equipment, or additional power source such as but notlimited to batteries, fuel cells, or the like. Optionally, the modulereplacing one or more of the modules 701-704 may provide storage foradditional disposables and/or reagents or fluids. It should beunderstood that although FIG. 7A shows only four modules mounted on thesupport structure, other embodiments having fewer or more modules arenot excluded from this horizontal mounting configuration. It should alsobe understood that configurations may also be run with not every bay orslot occupied by a module, particularly in any scenario wherein one ormore types of modules draw more power that other modules. In such aconfiguration, power otherwise directed to an empty bay can be used bythe module that may draw more power than the others.

In one non-limiting example, each module is secured to the supportstructure 732 with the aid of an attachment member. In an embodiment, anattachment member is a hook fastened to either the module or the bay. Insuch a case, the hook is configured to slide into a receptacle of eitherthe module or the bay. In another embodiment, an attachment memberincludes a fastener, such as a screw fastener. In another embodiment, anattachment member is formed of a magnetic material. In such a case, themodule and bay may include magnetic materials of opposite polarities soas to provide an attractive force to secure the module to the bay. Inanother embodiment, the attachment member includes one or more tracks orrails in the bay. In such a case, a module includes one or morestructures for mating with the one or more tracks or rails, therebysecuring the module to the support structure 732. Optionally, power maybe provided by the rails.

An example of a structure that may permit a module to mate with asupport structure 732 may include one or more pins. In some cases,modules receive power directly from the support structure 732. In somecases, a module may be a power source like a lithium ion, or fuel cellpowered battery that powers the device internally. In an example, themodules are configured to mate with the support structure 732 with theaid of rails, and power for the modules comes directly from the rails.In another example, the modules mate with the support structure 732 withthe aid of attachment members (rails, pins, hooks, fasteners), but poweris provided to the modules wirelessly, such as inductively (i.e.,inductive coupling).

Referring now to FIG. 7B, yet another embodiment of a system 740 isshown with a plurality of modules 701 to 706. FIG. 7B shows that asupport structure 742 is provided that can allow a transport device 744to move along the X, Y, and/or optionally Z axis to transport elementssuch as but not limited sample vessels, tips, cuvettes, or the likewithin a module and/or between modules. The transport device 744 can beconfigured to access either column of modules. Optionally, someembodiments may have more than one transport device 744 to providehigher throughput of transport capabilities for vessels or otherelements between modules. For clarity, the transport device 744 shown inphantom may represent a second transport device 744. Alternatively, itcan also be used to show where the transport device 744 is located whenservice the second column of modules. It should also be understood thatembodiments having still further rows and/or columns can also be createdby using a larger support structure to accommodate such a configuration.

It should be understood that, like the embodiment of FIG. 7, modules701-706 may all be modules that are identical to one another. In anotherembodiment, at least some of the modules are different from one another.In an example, the first, second, third, and/or fourth modules 701-706may be replaced by one or more other modules that can occupy thelocation of the module being replaced. The other modules may optionallyprovide different functionality such as but not limited to a replacingone of the modules 701-706 with one or more cytometry modules 707,communications modules, storage modules, sample preparation modules,slide preparation modules, tissue preparation modules, or the like.

It should be understood that although FIG. 7B shows only six modulesmounted on the support structure, other embodiments having fewer or moremodules are not excluded from this horizontal and vertical mountingconfiguration. It should also be understood that configurations may alsobe run with not every bay or slot occupied by a module, particularly inany scenario wherein one or more types of modules draw more power thatother modules. In such a configuration, power otherwise directed to anempty bay can be used by the module that may draw more power than theothers.

Referring now to FIG. 7C, yet another embodiment of a system 750 isshown with a plurality of modules 701, 702, 703, 704, 706, and 707. FIG.7C also shows that they system 750 has an additional module 752 that canwith one or more modules that provide a different hardware configurationsuch as but not limited to provide a thermal controlled storage chamberfor incubation, storage between testing, or storage after testing.Optionally, the module replacing one or more of the modules 701-704 canprovide a non-assay related functionality, such as but not limited toadditional telecommunication equipment for the system 730, additionalimaging or user interface equipment, or additional power source such asbut not limited to batteries, fuel cells, or the like. Optionally, themodule replacing one or more of the modules 701-707 may provide storagefor additional disposables and/or reagents or fluids.

It should be understood that although FIG. 7C shows seven modulesmounted on the support structure, other embodiments having fewer or moremodules are not excluded from this mounting configuration. It shouldalso be understood that configurations may also be run with not everybay or slot occupied by a module, particularly in any scenario whereinone or more types of modules draw more power that other modules. In sucha configuration, power otherwise directed to an empty bay can be used bythe module that may draw more power than the others.

In some embodiments, the modules 701-706 are in communication with oneanother and/or a controller of the system 700 by way of a communicationsbus (“bus”), which may include electronic circuitry and components forfacilitating communication among the modules and/or the controller. Thecommunications bus includes a subsystem that transfers data between themodules and/or controller of the system 700. A bus may bring variouscomponents of the system 700 in communication with a central processingunit (CPU), memory (e.g., internal memory, system cache) and storagelocation (e.g., hard disk) of the system 700.

A communications bus may include parallel electrical wires with multipleconnections, or any physical arrangement that provides logicalfunctionality as a parallel electrical bus. A communications bus mayinclude both parallel and bit-serial connections, and can be wired ineither a multidrop (i.e., electrical parallel) or daisy chain topology,or connected by switched hubs. In an embodiment, a communications busmay be a first generation bus, second generation bus or third generationbus. The communications bus permits communication between each of themodules and other modules and/or the controller. In some situations, thecommunications bus enables communication among a plurality of systems,such as a plurality of systems similar or identical to the system 700.

The system 700 may include one or more of a serial bus, parallel bus, orself-repairable bus. A bus may include a master scheduler that controldata traffic, such as traffic to and from modules (e.g., modules701-706), controller, and/or other systems. A bus may include anexternal bus, which connects external devices and systems to a mainsystem board (e.g., motherboard), and an internal bus, which connectsinternal components of a system to the system board. An internal busconnects internal components to one or more central processing units(CPUs) and internal memory.

In some embodiments, the communication bus may be a wireless bus. Thecommunications bus may be a Firewire (IEEE 1394), USB (1.0, 2.0, 3.0, orothers), or Thunderbolt.

In some embodiments, the system 700 includes one or more buses selectedfrom the group consisting of Media Bus, Computer Automated Measurementand Control (CAMAC) bus, industry standard architecture (ISA) bus, USBbus, Firewire, Thunderbolt, extended ISA (EISA) bus, low pin count bus,MBus, MicroChannel bus, Multibus, NuBus or IEEE 1196, OPTi local bus,peripheral component interconnect (PCI) bus, Parallel AdvancedTechnology Attachment (ATA) bus, Q-Bus, S-100 bus (or IEEE 696), SBus(or IEEE 1496), SS-50 bus, STEbus, STD bus (for STD-80 [8-bit] and STD32[16-/32-bit]), Unibus, VESA local bus, VMEbus, PC/104 bus, PC/104 Plusbus, PC/104 Express bus, PCI-104 bus, PCIe-104 bus, 1-Wire bus,HyperTransport bus, Inter-Integrated Circuit (I²C) bus, PCI Express (orPCIe) bus, Serial ATA (SATA) bus, Serial Peripheral Interface bus, UNI/Obus, SMBus, 2-wire or 3-wire interface, self-repairable elasticinterface buses and variants and/or combinations thereof.

In some situations, the system 700 includes a Serial PeripheralInterface (SPI), which is an interface between one or moremicroprocessors and peripheral elements or I/O components (e.g., modules701-706) of the system 700. The SPI can be used to attach 2 or more, or3 or more, or 4 or more, or 5 or more, or 6 or more, or 7 or more, or 8or more, or 9 or more, or 10 or more or 50 or more or 100 or more SPIcompatible I/O components to a microprocessor or a plurality ofmicroprocessors. In other instances, the system 700 includes RS-485 orother standards.

In an embodiment, an SPI is provided having an SPI bridge having aparallel and/or series topology. Such a bridge allows selection of oneof many SPI components on an SPI I/O bus without the proliferation ofchip selects. This is accomplished by the application of appropriatecontrol signals, described below, to allow daisy chaining the device orchip selects for the devices on the SPI bus. It does however retainparallel data paths so that there is no Daisy Chaining of data to betransferred between SPI components and a microprocessor.

In some embodiments, an SPI bridge component is provided between amicroprocessor and a plurality of SPI I/O components which are connectedin a parallel and/or series (or serial) topology. The SPI bridgecomponent enables parallel SPI using MISO and MOSI lines and serial(daisy chain) local chip select connection to other slaves (CSL/). In anembodiment, SPI bridge components provided herein resolve any issuesassociated with multiple chip selects for multiple slaves. In anotherembodiment, SPI bridge components provided herein support four, eight,sixteen, thirty two, sixty four or more individual chip selects for fourSPI enabled devices (CS1/−CS4/). In another embodiment, SPI bridgecomponents provided herein enable four times cascading with externaladdress line setting (ADR0-ADR1). In some situations, SPI bridgecomponents provided herein provide the ability to control up to eight,sixteen, thirty two, sixty four or more general output bits for controlor data. SPI bridge components provided herein in some cases enable thecontrol of up to eight, sixteen, thirty two, sixty four or more generalinput bits for control or data, and may be used for deviceidentification to the master and/or diagnostics communication to themaster.

FIG. 41A shows an SPI bridge scheme having master and parallel-seriesSPI slave bridges, in accordance with an embodiment of the invention.The SPI bus is augmented by the addition of a local chip select (CSL/),module select (MOD_SEL) and select data in (DIN_SEL) into a SPI bridgeto allow the addition of various system features, including essentialand non-essential system features, such as cascading of multiple slavedevices, virtual daisy chaining of device chip selects to keep themodule-to-module signal count at an acceptable level, the support formodule identification and diagnostics, and communication to non-SPIelements on modules while maintaining compatibility with embedded SPIcomplaint slave components. FIG. 41B shows an example of an SPI bridge,in accordance with an embodiment of the invention. The SPI bridgeincludes internal SPI control logic, a control register (8 bit, asshown), and various input and output pins.

Each slave bridge is connected to a master (also “SPI master” and“master bridge” herein) in a parallel-series configuration. The MOSI pinof each slave bridge is connected to the MOSI pin of the master bridge,and the MOSI pins of the slave bridges are connected to one another.Similarly, the MISO pin of each slave bridge is connected to the MISOpin of the master bridge, and the MISO pins of the slave bridges areconnected to one another.

Each slave bridge may be a module (e.g., one of the modules 701-706 ofFIG. 7) or a component in a module. In an example, the First SlaveBridge is the first module 701, the Second Slave Bridge is the secondmodule 702, and so on. In another example, the First Slave Bridge is acomponent (e.g., one of the components 910 of FIG. 9) of a module.

FIG. 41C shows a module component diagram with interconnected modulepins and various components of a master bridge and slave bridge, inaccordance with an embodiment of the invention. FIG. 41D shows slavebridges connected to a master bridge, in accordance with an embodimentof the invention. The MISO pin of each slave bridge is in electricalcommunication with a MOSI pin of the master bridge. The MOSI pin of eachslave bridge is in electrical communication with a MISO pin of themaster bridge. The DIN_SEL pin of the first slave bridge (left) is inelectrical communication with the MOSI pin of the first slave bridge.The DOUT_SEL pin of the first slave bridge is in electricalcommunication with the DIN_SEL of the second slave (right). Additionalslave bridges may be connected as the second slave by bringing theDIN_SEL pins of each additional slave bridge in electrical communicationwith a DOUT_SEL pin of a previous slave bridge. In such fashion, theslave bridge are connected in a parallel-series configuration.

In some embodiments, CLK pulses directed to connected SPI-Bridgescapture the state of DIN_SEL Bits shifted into the Bridges at theassertion of the Module Select Line (MOD_SEL). The number of DIN_SELbits corresponds to the number of modules connected together on aparallel-series SPI-Link. In an example, if the two modules areconnected in a parallel-series configuration (e.g. RS486), the number ofDIN_SEL is equal to two.

In an embodiment, SPI-Bridges which latch a ‘1’ during the moduleselection sequence become the ‘selected module’ set to receive 8 bitcontrol word during a following element selection sequence. EachSPI-Bridge may access up to 4 cascaded SPI Slave devices. Additionally,each SPI-Bridge may have an 8-Bit GP Receive port and 8-Bit GP TransmitPort. An ‘element selection’ sequence writes an 8 bit word into the‘selected module’ SPI-Bridge control register to enable subsequenttransactions with specific SPI devices or to read or write data via theSPI-Bridge GPIO port.

In an embodiment, element selection takes place by assertion of thelocal chip select line (CSL/) then clocking the first byte of MOSItransferred data word into the control register. In some cases, theformat of the control register is CS4 CS3 CS2 CS1 AD1 AD0 R/W N. Inanother embodiment, the second byte is transmit or receive data. WhenCSL/ is de-asserted, the cycle is complete.

In an SPI transaction, following the element selection sequence,subsequent SPI slave data transactions commence. The SPI CS/ (which maybe referred to as SS/) is routed to one of 4 possible bridged devices,per the true state of either CS4, CS3, CS2 or CS1. Jumper bits AD0, AD1are compared to AD0, AD1 of the control register allow up to fourSPI-Bridges on a module.

FIG. 41E shows a device having a plurality of modules mounted on a SPIlink of a communications bus of the device, in accordance with anembodiment of the invention. Three modules are illustrated, namelyModule 1, Module 2 and Module 3. Each module includes one or more SPIbridges for bringing various components of a module in electricalconnection with the SPI link, including a master controller (includingone or more CPU's) in electrical communication with the SPI link. Module1 includes a plurality of SPI slaves in electrical communication witheach of SPI Bridge 00, SPI Bridge 01, SPI Bridge 10 and SPI Bridge 11.In addition, each module includes a Receive Data controller, TransmitData controller and Module ID jumpers.

In other embodiments, the modules 701-706 are configured to communicatewith one another and/or one or more controllers of the system 700 withthe aid of a wireless communications bus (or interface). In an example,the modules 701-706 communicate with one another with the aid of awireless communications interface. In another example, one or more ofthe modules 701-706 communicate with a controller of the system 700 withthe aid of a wireless communications bus. In some cases, communicationamong the modules 701-706 and/or one or more controllers of the systemis solely by way of a wireless communications bus. This mayadvantageously preclude the need for wired interfaces in the bays foraccepting the modules 701-706. In other cases, the system 700 includes awired interface that works in conjunction with a wireless interface ofthe system 700.

Although the system 700, as illustrated, has a single rack, a system,such as the system 700, may have multiple racks. In some embodiments, asystem has at most 1, or 2, or 3, or 4, or 5, or 6, or 7, or 8, or 9, or10, or 20, or 30, or 40, or 50, or 100, or 1000, or 10,000 racks. In anembodiment, the system has a plurality of racks disposed in aside-by-side configuration.

FIG. 8 shows an example of a multi-rack system. For example, a firstrack 800 a may be connected and/or adjacent to a second rack 800 b. Eachrack may include one or more module 810. In another embodiment, thesystem includes a plurality of racks that are disposed vertically inrelation to one another—that is, one rack on top of another rack. Insome embodiments, the racks may form a vertical array (e.g., one or moreracks high and one or more racks wide), a horizontal array (one or moreracks wide, one or more racks long), or a three-dimensional array (oneor more racks high, one or more racks wide, and one or more racks long).

In some embodiments, the modules may be disposed on the racks, dependingon rack configuration. For example, if vertically oriented racks areplaced adjacent to one another, modules may be disposed vertically alongthe racks. If horizontally oriented racks are placed on top of oneanother, modules may be disposed horizontally along the racks. Racks maybe connected to one another via any sort of connecting interface,including those previously described for modules. Racks may or may notcontact one another. Racks may be mechanically and/or electricallyconnected to one another.

In another embodiment, the system includes a plurality of racks, andeach rack among the plurality of racks is configured for a differentuse, such as sample processing. In an example, a first rack isconfigured for sample preparation and cytometry and a second rack isconfigured for sample preparation and agglutination. In anotherembodiment, the racks are disposed horizontally (i.e., along an axisorthogonal to the gravitational acceleration vector). In anotherembodiment, the system includes a plurality of racks, and two or moreracks among the plurality of racks are configured for the same use, suchas sample preparation or processing.

In some cases, a system having a plurality of racks includes a singlecontroller that is configured to direct (or facilitate) sampleprocessing in each rack. In other cases, each individual rack among aplurality of racks includes a controller configured to facilitate sampleprocessing in the individual rack. The controllers may be in network orelectrical communication with one another.

A system having a plurality of racks may include a communications bus(or interface) for bringing the plurality of racks in communication withone another. This permits parallel processing among the racks. Forinstance, for a system including two racks commutatively coupled to oneanother with the aid of a communications bus, the system processes afirst sample in a first of the two racks while the system processes asecond sample in a second of the two racks.

A system having a plurality of racks may include one or more samplehandling systems for transferring samples to and from racks. In anexample, a system includes three racks and two sample handling systemsto transfer samples to and from each of the first, second and thirdracks.

In some embodiments, sample handling systems are robots or robotic-armsfor facilitating sample transfer among racks, among modules in a rack,and/or within modules. In some embodiments, each module may have one ormore robots. The robots may be useful for moving components within oramongst different modules or other components of a system. In otherembodiments, sample handling systems are actuator (e.g., electricalmotors, pneumatic actuators, hydraulic actuators, linear actuators, combdrive, piezoelectric actuators and amplified piezoelectric actuators,thermal bimorphs, micromirror devices and electroactive polymers)devices for facilitating sample transfer among racks or modules in arack. In other embodiments, sample handling systems include pipettes,such as positive displacement, suction-type or air displacement pipetteswhich may optionally have robotic capabilities or robots with pipettingcapability. One or more robots may be useful for transferring samplingsystems from one location to another.

The robotic arm (also “arm” here) is configured to transfer (or shuttle)samples to and from modules or, in some cases, among racks. In anexample, an arm transfers samples among a plurality of verticallyoriented modules in a rack. In another example, an arm transfers samplesamong a plurality of horizontally oriented modules in a rack. In anotherexample, an arm transfers samples among a plurality of horizontally andvertically oriented modules in a rack.

Each arm may include a sample manipulation device (or member) forsupporting a sample during transport to and from a module and/or one ormore other racks. In an embodiment, the sample manipulation device isconfigured to support a tip or vessel (e.g., container, vial) having thesample. The sample manipulation device may be configured to support asample support, such as a microcard or a cartridge. Alternatively, themanipulation device may have one or more features that may permit themanipulation device to serve as a sample support. The samplemanipulation device may or may not include a platform, gripper, magnet,fastener, or any other mechanism that may be useful for the transport.

In some embodiments, the arm is configured to transfer a module from onebay to another. In an example, the arm transfers a module from a firstbay in a first rack to a first bay in a second rack, or from the firstbay in the first rack to a second bay in the second rack.

The arm may have one or more actuation mechanism that may permit the armto transfer the sample and/or module. For example, one or more motor maybe provided that may permit movement of the arm.

In some instances, the arm may move along a track. For example, avertical and/or horizontal track may be provided. In some instances, therobot arm may be a magnetic mount with a kinematic locking mount.

In some embodiments, robots, such as a robotic arm, may be providedwithin a device housing. The robots may be provided within a rack,and/or within a module. Alternatively, they may be external to a rackand/or module. They may permit movement of components within a device,between tracks, between modules, or within modules. The robots may moveone or more component, including but not limited to a sample handlingsystem, such as a pipette, vessel/tip, cartridge, centrifuge, cytometer,camera, detection unit, thermal control unit, assay station or system,or any other component described elsewhere herein. The components may bemovable within a module, within a rack, or within the device. Thecomponents may be movable within the device even if no rack or module isprovided within the device. The robots may move one or more module. Themodules may be movable within the device. The robots may move one ormore racks. The racks may be movable within the device.

The robots may move using one or more different actuation mechanism.Such actuation mechanisms may use mechanical components,electromagnetic, magnetism, thermal properties, piezoelectricproperties, optics, or any other properties or combinations thereof. Forexample, the actuation mechanisms may use a motor (e.g., linear motor,stepper motor), lead screw, magnetic track, or any other actuationmechanism. In some instances, the robots may be electronically,magnetically, thermally or optically controlled.

FIG. 68A provides an example of a magnetic way of controlling theposition of a robot or other item. A top view shows an array of magnets6800. A coil support structure 6810 may be provided adjacent to themagnets. A coil support structure may be made from electricallyconductive, weak magnetic material.

The array of magnets may include a strip of magnets, or an m×n array ofmagnets, where m and/or n is greater than or equal to 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 40, 50, or100.

FIG. 68B provides a side view of the magnetic control. A coil supportstructure 6810 may have one, two, three, four, five, six, seven, eightor more conducting coils 6820 thereon. The coil support structure may beadjacent to an array of magnets 6800.

Passive damping may be provided as well as use of electricallyconductive magnetic materials.

The actuation mechanisms may be capable of moving with very highprecision. For example, the robots may be capable of moving with aprecision of within about 0.01 nm, 0.05 nm, 0.1 nm, 0.5 nm, 1 nm, 5 nm,10 nm, 30 nm, 50 nm, 75 nm, 100 nm, 150 nm, 200 nm, 250 nm, 300 nm, 400nm, 500 nm, 600 nm, 700 nm, 800 nm, 900 nm, 1 μm, 1.5 μm, 2 μm, 3 μm, 4μm, 5 μm, 7 μm, 10 μm, 15 μm, 20 μm, 25 μm, 30 μm, 40 μm, 50 μm, 75 μm,100 μm, 150 μm, 200 μm, 250 μm, 300 μm, 500 μm, 750 μm, 1 mm, 2 mm, or 3mm.

The robots may be capable of moving in any direction. The robots may becapable of moving in a lateral direction (e.g., horizontal direction)and/or a vertical direction. A robot may be capable of moving within ahorizontal plane, and/or a vertical plane. A robot may be capable ofmoving in an x, y, and/or z direction wherein an x-axis, y-axis, andz-axis are orthogonal to one another. Some robots may only move withinone dimension, two dimensions, and/or three dimensions.

In some situations, the term “system” as used herein may refer to a“device” or “sample processing device” disclosed herein, unless thecontext clearly dictates otherwise.

Plug-and-Play

In an aspect of the invention, plug-and-play systems are described. Theplug-and-play systems are configured to assay at least one sample, suchas a tissue or fluid sample, from a subject.

In some embodiments, the plug-and-play system comprises a supportingstructure having a mounting station configured to support a module amonga plurality of modules. The module is detachable from the mountingstation. In some cases, the module is removably detachable—that is, themodule may be removed from the mounting station and returned to itsoriginal position on the mounting station. Alternatively, the module maybe replaced with another module.

In an embodiment, the module is configured to perform without the aid ofanother module in the system (a) at least one sample preparationprocedure selected from the group consisting of sample processing,centrifugation, magnetic separation, or (b) at least one type of assayselected from the group consisting of immunoassay, nucleic acid assay,receptor-based assay, cytometric assay, colorimetric assay, enzymaticassay, electrophoretic assay, electrochemical assay, spectroscopicassay, chromatographic assay, microscopic assay, topographic assay,calorimetric assay, turbidimetric assay, agglutination assay,radioisotope assay, viscometric assay, coagulation assay, clotting timeassay, protein synthesis assay, histological assay, culture assay,osmolarity assay, and/or other types of assays or combinations thereof.

In an embodiment, the module is configured to be in electrical,electro-magnetic or optoelectronic communication with a controller. Thecontroller is configured to provide one or more instructions to themodule or individual modules of the plurality of modules to facilitateperformance of the at least one sample preparation procedure or the atleast one type of assay.

In an embodiment, the system is in communication with a controller forcoordinating or facilitating the processing of samples. In anembodiment, the controller is part of the system. In another embodiment,the controller is remotely located with respect to the system. In anexample, the controller is in network communication with the system.

In an embodiment, a module is coupled to a support structure. Thesupport structure may be a rack having a plurality of bays for acceptinga plurality of modules. The support structure is part of the systemconfigured to accept the module. In an embodiment, the module ishot-swappable—that is, the module may be exchanged with another moduleor removed from the support structure while the system is processingother samples.

In some embodiments, upon a user hot-swapping a first module for asecond module, the system is able to detect and identify the secondmodule and update a list of modules available for use by the system.This permits the system to determine which resources are available foruse by the system for processing a sample. For instance, if a cytometrymodule is swapped for an agglutination module and the system has noother cytometry modules, then the system will know that the system isunable to perform cytometry on a sample.

The plurality of modules may include the same module or differentmodules. In some cases, the plurality of modules are multi-purpose (ormulti-use) modules configured for various preparation and/or processingfunctionalities. In other cases, the plurality of modules may bespecial-use (or special-purpose) modules configured for fewerfunctionalities than the multi-purpose modules. In an example, one ormore of the modules is a special-use module configured for cytometry.

In some embodiments, the system is configured to detect the type ofmodule without the need for any user input. Such plug-and-playfunctionality advantageously enables a user to insert a module into thesystem for use without having to input any commands or instructions.

In some situations, the controller is configured to detect a module. Insuch a case, when a user plugs a module into the system, the systemdetects the module and determines whether the module is a multi-usemodule or special-use module. In some cases, the system is able todetect a module with the use of an electronic identifier, which mayinclude a unique identifier. In other cases, the system is able todetect the module with the aid of a physical identifier, such as a barcode or an electronic component configured to provide a unique radiofrequency identification (RFID) code, such as an RFID number or a uniqueID through the system bus.

The system may detect a module automatically or upon request from a useror another system or electronic component in communication with thesystem. In an example, upon a user inputting the module 701 into thesystem 700, the system 700 detects the module, which may permit thesystem 700 to determine the type of module (e.g., cytometry module).

In some situations, the system is configured to also determine thelocation of the module, which may permit the system to build a virtualmap of modules, such as, e.g., for facilitating parallel processing (seebelow). In an example, the system 700 is configured to detect thephysical location of each of the modules 701-706. In such a case, thesystem 700 knows that the first module 701 is located in a first port(or bay) of the system 700.

Modules may have the same component or different components. In anembodiment, each module has the same components, such as those describedabove in the context of FIG. 7. That is, each module includes pipettesand various sample processing stations. In another embodiment, themodules have different components. In an example, some modules areconfigured for cytometry assays while other are configured foragglutination assays.

In another embodiment, a shared module may be a dedicated cooling orheating unit that is providing cooling or heating capabilities to thedevice or other modules as needed.

In another embodiment, a shared resource module may be a rechargeablebattery pack. Examples of batteries may include, but are not limited to,zinc-carbon, zinc-chloride, alkaline, oxy-nickel hydroxide, lithium,mercury oxide, zinc-air, silver oxide, NiCd, lead acid, NiMH, NiZn, orlithium ion. These batteries may be hot-swappable or not. Therechargeable battery may be coupled with external power source. Therechargeable battery module may be recharged while the device is pluggedinto an external power source or the battery module may be taken out ofdevice and recharged externally to the device in a dedicated rechargingstation or directly plugged into an external power supply. The dedicatedrecharging station may be the device or be operatively connected to thedevice (e.g., recharging can be done via induction without directphysical contact). The recharging station may be a solar poweredrecharging station or may be powered by other clean or conventionalsources. The recharging station may be powered by a conventional powergenerator. The battery module may provide Uninterrupted Power Supply(UPS) to the device or bank of devices in case of power interruptionsfrom external supply.

In another embodiment, the shared resource module may be a ‘computefarm’ or collection of high performance general purpose or specificpurpose processors packed together with appropriate cooling as a modulededicated to high performance computing inside the device or to beshared by collection of devices.

In another embodiment, a module may be an assembly of high performanceand/or high capacity storage devices to provide large volume of storagespace (e.g. 1 TB, 2 TB, 10 TB, 100 TB, 1 PB, 100 PB or more) on thedevice to be shared by all modules, modules in other devices that may besharing resources with the device and even by the external controller tocache large amounts of data locally to a device or a physical site orcollection of sites or any other grouping of devices.

In another embodiment, a shared module may be a satellite communicationmodule that is capable of providing communication capabilities tocommunicate with satellite from the device or other devices that may besharing resources.

In another embodiment, the module may be an internet router and/or awireless router providing full routing and/or a hotspot capability tothe device or bank of devices that are allowed to share the resources ofthe device.

In some embodiments, the module, alone or in combination with othermodules (or systems) provided herein, may act as a ‘data center’ foreither the device or bank of devices allowed to share the resources ofthe device providing high performance computing, high volume storage,high performance networking, satellite or other forms of dedicatedcommunication capabilities in the device for a given location or site orfor multiple locations or sites.

In one embodiment, a shared module may be a recharging station forwireless or wired peripherals that are used in conjunction with thedevice.

In one embodiment, a shared module may be a small refrigeration ortemperature control storage unit to stores, samples, cartridges, othersupplies for the device.

In another embodiment, a module may be configured to automaticallydispense prescription or other pharmaceutical drugs. The module may alsohave other components such as packet sealers and label printers thatmake packaging and dispensing drugs safe and effective. The module maybe programmed remotely or in the device to automatically dispense drugsbased on real time diagnosis of biological sample, or any otheralgorithm or method that determines such need. The system may have theanalytics for pharmacy decision support to support the module aroundtreatment decisions, dosing, and other pharmacy-related decisionsupport.

Modules may have swappable components. In an example, a module has apositive displacement pipette that is swappable with the same type ofpipette or a different type of pipette, such as a suction-type pipette.In another example, a module has an assay station that is swappable withthe same type of assay station (e.g., cytometry) or a different type ofassay station (e.g., agglutination). The module and system areconfigured to recognize the modules and components in the modules andupdate or modify processing routines, such as parallel processingroutines, in view of the modules coupled to the system and thecomponents in each of the modules.

In some cases, the modules may be external to the device and connectedto the device through device's bus (e.g. via a USB port).

FIG. 9 shows an example of a module 900 having one or more components910. A module may have one or more controller. The components 910 areelectrically coupled to one another and/or the controller via acommunications bus (“Bus”), such as, for example, a bus as describedabove in the context of FIG. 7. In an example, the module 900 includes aone or more buses selected from the group consisting of Media Bus,Computer Automated Measurement and Control (CAMAC) bus, industrystandard architecture (ISA) bus, extended ISA (EISA) bus, low pin countbus, MBus, MicroChannel bus, Multibus, NuBus or IEEE 1196, OPTi localbus, peripheral component interconnect (PCI) bus, Parallel AdvancedTechnology Attachment (ATA) bus, Q-Bus, S-100 bus (or IEEE 696), SBus(or IEEE 1496), SS-50 bus, STEbus, STD bus (for STD-80 [8-bit] and STD32[16-/32-bit]), Unibus, VESA local bus, VMEbus, PC/104 bus, PC/104 Plusbus, PC/104 Express bus, PCI-104 bus, PCIe-104 bus, 1-Wire bus,HyperTransport bus, Inter-Integrated Circuit (I²C) bus, PCI Express (orPCIe) bus, Serial ATA (SATA) bus, Serial Peripheral Interface bus, UNI/Obus, SMBus, self-repairable elastic interface buses and variants and/orcombinations thereof. In an embodiment, the communications bus isconfigured to communicatively couple the components 910 to one anotherand the controller. In another embodiment, the communications bus isconfigured to communicatively couple the components 910 to thecontroller. In an embodiment, the communications bus is configured tocommunicatively couple the components 910 to one another. In someembodiments, the module 900 includes a power bus that provides power toone or more of the components 910. The power bus may be separate fromthe communications bus. In other embodiments, power is provided to oneor more of the components with the aid of the communications bus.

In an embodiment, the components 910 may be swappable, such ashot-swappable. In another embodiment, the components 910 are removablefrom the module 900. The components 910 are configured for samplepreparation, processing and testing. Each of the components 910 may beconfigured to process a sample with the aid of one or more sampleprocessing, preparation and/or testing routines.

In the illustrated example, the module 900 includes six components 910:a first component (Component 1), second component (Component 2), thirdcomponent (Component 3), fourth component (Component 4), fifth component(Component 5), and sixth component (Component 6). The module 900generally includes 1 or more, or 2 or more, or 3 or more, or 4 or more,or 5 or more, or 6 or more, or 7 or more, or 8 or more, or 9 or more, or10 or more, or 20 or more, or 30 or more, or 40 or more, or 50 or more,or 100 or more components 910. The components 910, with the aid of thecontroller communicative (and electrically) coupled to the components910, are configured for serial and/or parallel processing of a sample.

In an example, Component 1 is a centrifuge, Component 2 is aspectrophotometer, Component 3 is a Nucleic Acid (assay station andComponent 4 is a PMT station, Component 5 is a tip holder and Component6 is a sample washing station.

In an embodiment, the components are configured to process a sample inseries. In such a case, a sample is processed in the components insequence (i.e., Component 1, Component 2, etc.). In another embodiment,sample processing is not necessarily sequential. In an example, a sampleis first processed in Component 4 followed by Component 1.

In an embodiment, the components 910 process samples in parallel. Thatis, a component may process a sample while one or more other componentsprocess the sample or a different sample. In an example, Component 1processes a sample while Component 2 processes a sample. In anotherembodiment, the components 910 process sample sequentially. That is,while one component processes a sample, another component does notprocess a sample.

In some embodiments, the module 900 includes a sample handling systemconfigured to transfer a sample to and from the components 910. In anembodiment, the sample handling system is a positive displacementpipette. In another embodiment, the sample handling system is asuction-type pipette. In another embodiment, the sample handling systemis an air-displacement pipette. In another embodiment, the samplehanding system includes one or more of a suction-type pipette, positivedisplacement pipette and air-displacement pipette. In anotherembodiment, the sample handing system includes any two of a suction-typepipette, positive displacement pipette and air-displacement pipette. Inanother embodiment, the sample handing system includes a suction-typepipette, positive displacement pipette and air-displacement pipette.

The components 910 may be connected via bus architectures providedherein. In an example, the components 910 are connected via theparallel-series configuration described in the context of FIGS. 41A-41E.That is, each component 910 may be connected to an SPI slave bridge thatis in turn connected to a master bridge. In other embodiments, thecomponents 910 are connected in a series (or daisy-chain) configuration.In other embodiments, the components 910 are connected in a parallelconfiguration.

In some embodiments, the components 910 are swappable with othercomponents. In an embodiment, each component is swappable with the samecomponent (i.e., another component having the same functionality). Inanother embodiment, each component is swappable with a differentcomponent (i.e., a component having different functionality). Thecomponents 910 are hot swappable or removable upon shutdown of themodule 900.

FIG. 10 shows a system 1000 having a plurality of modules mounted tobays of the system 1000, in accordance with an embodiment of theinvention. The system includes a first module (Module 1), second module(Module 2) and third module (Module 3). The system 1000 includes acommunications bus (“Bus”) for bringing a controller of the system 1000in communication with each of the modules. The communications bus (also“system bus” herein) of the system 1000 is also configured to bring themodules in communication with one another. In some situations, thecontroller of the system 1000 is optional.

With continued reference to FIG. 10, each module includes a plurality ofstations (or sub-modules), designated by Mxy, wherein ‘x’ designates themodule and ‘y’ designates the station. Each module optionally includes acontroller that is communicatively coupled to each of the stations via acommunications bus (also “module bus” herein). In some cases, acontroller is communicatively coupled to the system bus through themodule bus.

Module 1 includes a first station (M11), second station (M12), thirdstation (M13) and controller (C1). Module 2 includes a first station(M21), second station (M22), third station (M23) and controller (C2).Module 3 includes a first station (M31) and controller (C3). Thecontrollers of the modules are communicatively coupled to each of thestations via a communications bus. The stations are selected from thegroup consisting of preparation stations, assaying stations anddetection stations. Preparation stations are configured for samplepreparation; assaying stations are configured for sample assaying; anddetection stations are configured for analyte detection.

In an embodiment, each module bus is configured to permit a station tobe removed such that the module may function without the removedstation. In an example, M11 may be removed from module 1 whilepermitting M12 and M13 to function. In another embodiment, each stationis hot-swappable with another station—that is, one station may bereplaced with another station without removing the module or shuttingdown the system 1000.

In some embodiments, the stations are removable from the modules. Inother embodiments, the stations are replaceable by other stations. In anexample, M11 is replaced by M22.

With respect to a particular module, each station may be different ortwo or more stations may be the same. In an example, M11 is a centrifugeand M12 is an agglutination station. As another example, M22 is anucleic acid assay station and M23 is an x-ray photoelectronspectroscopy station.

Two or more of the modules may have the same configuration of stationsor a different configuration. In some situations, a module may be aspecialized module. In the illustrated embodiment of FIG. 10, module 3has a single station, M31, that is communicatively coupled to C3.

The system 1000 includes a sample handling system for transferringsamples to and from the modules. The sample handling system includes apositive displacement pipette, suction-type pipette and/orair-displacement pipette. The sample handling system is controlled bythe controller of the system 1000. In some situations, the samplehandling system is swappable by another sample handling system, such asa sample handling system specialized for certain uses.

With continued reference to FIG. 10, each module includes a samplehandling system for transferring samples to and from the stations. Thesample handling system includes a positive displacement pipette,suction-type pipette and/or air-displacement pipette. The samplehandling system is controlled by a controller in the module.Alternatively, the sample handling system is controlled by thecontroller of the system 1000.

Parallel Processing and Dynamic Resource Sharing

In another aspect of the invention, methods for processing a sample areprovided. The methods are used to prepare a sample and/or perform one ormore sample assays.

In some embodiments, a method for processing a sample comprisesproviding a system having plurality of modules as described herein. Themodules of the system are configured to perform simultaneously (a) atleast one sample preparation procedure selected from the groupconsisting of sample processing, centrifugation, magnetic separation andchemical processing, and/or (b) at least one type of assay selected fromthe group consisting of immunoassay, nucleic acid assay, receptor-basedassay, cytometric assay, colorimetric assay, enzymatic assay,electrophoretic assay, electrochemical assay, spectroscopic assay,chromatographic assay, microscopic assay, topographic assay,calorimetric assay, turbidimetric assay, agglutination assay,radioisotope assay, viscometric assay, coagulation assay, clotting timeassay, protein synthesis assay, histological assay, culture assay,osmolarity assay, and/or other types of assays or combinations thereof.Next, the system tests for the unavailability of resources or thepresence of a malfunction of (a) the at least one sample preparationprocedure or (b) the at least one type of assay. Upon detection of amalfunction within at least one module, the system uses another moduleof the system or another system in communication with the system toperform the at least one sample preparation procedure or the at leastone type of assay.

In some embodiments, the system 700 of FIG. 7 is configured to allocateresource sharing to facilitate sample preparation, processing andtesting. In an example, one of the modules 701-706 is configured toperform a first sample preparation procedure while another of themodules 701-706 is configured to perform a second sample preparationprocedure that is different from the first sample preparation procedure.This enables the system 700 to process a first sample in the firstmodule 701 while the system 700 processes a second sample or a portionof the first sample. This advantageously reduces or eliminates downtime(or dead time) among modules in cases in which processing routines inmodules (or components within modules) require different periods of timeto reach completion. Even if processing routines reach completion withinthe same period of time, in situations in which the periods do notoverlap, parallel processing enables the system to optimize systemresources in cases. This may be applicable in cases in which a module isput to use after another module or if one module has a start time thatis different from that of another module.

The system 700 includes various devices and apparatuses for facilitatingsample transfer, preparation and testing. The sample handling system 708enables the transfer of a sample to and from each of the modules701-706. The sample handling system 708 may enable a sample to beprocessed in one module while a portion of the sample or a differentsample is transferred to or from another module.

In some situations, the system 700 is configured to detect each of themodules 701-706 and determine whether a bay configured to accept modulesis empty or occupied by a module. In an embodiment, the system 700 isable to determine whether a bay of the system 700 is occupied by ageneral or multi-purpose module, such as a module configured to performa plurality of tests, or a specialized module, such as a moduleconfigured to perform select tests. In another embodiment, the system700 is able to determine whether a bay or module in the bay is defectiveor malfunctioning. The system may then use other modules to performsample processing or testing.

A “multi-purpose module” is configured for a wide array of uses, such assample preparation and processing. A multi-purpose module may beconfigured for at least 2, or 3, or 4, or 5, or 6, or 7, or 8, or 9, or10, or 20, or 30, or 40, or 50 uses. A “special-use module” is a modulethat is configured for one or more select uses or a subset of uses, suchas at most 1, or 2, or 3, or 4, or 5, or 6, or 7, or 8, or 9, or 10, or20, or 30, or 50 uses. Such uses may include sample preparation,processing and/or testing (e.g., assay). A module may be a multi-purposemodule or special-use module.

In some cases, a special-use module may include sample preparationprocedures and/or tests not include in other modules. Alternatively, aspecial-use module includes a subset of sample preparation proceduresand/or tests included in other modules.

In the illustrated example of FIG. 7, the module 706 may be aspecial-use module. Special uses may include, for example, one or moreassays selected from cytometry, agglutination, microscopy and/or anyother assay described elsewhere herein.

In an example, a module is configured to perform cytometry only. Themodule is configured for use by the system 700 to perform cytometry. Thecytometry module may be configured to prepare and/or process a sampleprior to performing cytometry on the sample.

In some embodiments, systems are provided that are configured to processmultiple samples in parallel. The samples may be different samples orportions of the same sample (e.g., portions of a blood sample). Parallelprocessing enables the system to make use of system resources at timeswhen such resources would otherwise not be used. In such fashion, thesystem is configured to minimize or eliminate dead time betweenprocessing routines, such as preparation and/or assay routines. In anexample, the system assays (e.g., by way of cytometry) a first sample ina first module while the system centrifuges the same or a differentsample in a different module.

In some situations, the system is configured to process a first samplein a first component of a first module while the system processes asecond sample in a second component of the first module. The firstsample and second sample may be portions of a larger quantity of asample, such as portions of a blood sample, or different sample, such asa blood sample from a first subject and a blood sample from a secondsubject, or a urine sample from the first subject and a blood samplefrom the first subject. In an example, the system assays a first samplein the first module while the system centrifuges a second sample in thefirst module.

FIG. 11 shows a plurality of plots illustrating a parallel processingroutine, in accordance with an embodiment of the invention. Each plotillustrates processing in an individual module as a function of time(abscissa, or “x axis”). In each module, a step increase with timecorresponds to the start of processing and a step decrease with timecorresponds to the termination (or completion) of processing. The topplot shows processing in a first module, the middle plot showsprocessing in a second module, and the bottom plot shows processing in athird module. The first, second and third modules are part of the samesystem (e.g., system 700 of FIG. 7). Alternatively, the first, secondand/or third modules may be part of separate systems.

In the illustrated example, when the first module processes a firstsample, the second module processes a second sample and the third moduleprocesses a third sample. The first and third modules start processingat the same time, but processing times are different. This may be thecase if, for example, the first module processes a sample with the aidof an assay or preparation routine that is different from that of thethird module (e.g., centrifugation in the first module and cytometry inthe third module). Additionally, the first module takes twice as long tocomplete. In that time period, the third module processes a secondsample.

The second module starts processing a sample at a time that is laterthan the start time of the first and third modules. This may be the caseif, for example, the second module requires a period for completion ofsample processing that is different from that of the first and thirdmodules, or if the second module experiences a malfunction.

The modules may have the same dimensions (e.g., length, width, height)or different dimensions. In an example, a general or special-use modulehas a length, width and/or height that is different from that of anothergeneral or special-use module.

In some situations, systems and modules for processing biologicalsamples are configured to communicate with other systems to facilitatesample processing (e.g., preparation, assaying). In an embodiment, asystem communicates with another system by way of a wirelesscommunication interface, such as, e.g., a wireless network router,Bluetooth, radiofrequency (RF), opto-electronic, or other wireless modesof communication. In another embodiment, a system communicates withanother system by way of a wired communication, such as a wired network(e.g., the Internet or an intranet).

In some embodiments, point of service devices in a predetermined areacommunicate with one another to facilitate network connectivity, such asconnectivity to the Internet or an intranet. In some cases, a pluralityof point of service devices communicate with one another with the aid ofan intranet, such as an intranet established by one of the plurality ofpoint of service devices. This may permit a subset of a plurality ofpoint of service devices to connect to a network without a direct (e.g.,wired, wireless) network connection—the subset of the plurality of pointof service devices connect to the network with the aid of the networkconnectivity of a point of service device connected to the network. Withthe aid of such shared connectivity, one point of service device mayretrieve data (e.g., software, data files) without having to connect toa network. For instance, a first point of service device not connectedto a wide-area network may retrieve a software update by forming alocal-area connection or a peer-to-peer connection to a second point ofservice device. The first point of service device may then connect tothe wide-area network (or cloud) with the aid of the networkconnectivity of the second point of service device. Alternatively, thefirst point of service device may retrieve a copy of the software updatedirectly from the second point of service device.

In an example of shared connectivity, a first point of service devicesconnects (e.g., wireless connection) to a second point of servicedevice. The second point of service device is connected to a networkwith the aid of a network interface of the second point of servicedevice. The first point of service device may connect to the networkthrough the network connection of the second point of service device.

Log-Based Journaling and Fault Recovery

Another aspect of the invention provides methods for enabling devicesand systems, such as point of service devices, to maintain transactionrecords and/or operational log journals. Such methods enable systems anddevices provided herein, for example, to recover from a fault condition.

In some situations, point of service devices and modules haveoperational states that characterize the state of operation of suchdevices, such as, for example, sample centrifugation, sample transferfrom a first component to a second component, or nucleic acidamplification. In an embodiment, the operational state is a separate (ordiscrete) condition of a state of operation of a point of servicedevice.

Operational state may capture operations at various levels, such as atthe device level or system level. In an example, an operational stateincludes using a device (e.g., pipette). In another example, anoperational state includes moving a component of the device (e.g.,moving the pipette two inches to the left).

In some embodiments, a point of service device has a processing catalog(or operational catalog) having one or more operational matrices. Eachof the one or more matrices has discrete operational states of the pointof service system (or device) or one or more modules of the system. Theprocessing catalog may be generated by the point of service system ordevice, or another system on or associated with the point of servicesystem or device. In an example, the processing catalog is generatedupon initial system start or setup. In another example, the processingcatalog is generated upon request by a user or other system, such as amaintenance system.

In an embodiment, a point of service system generates a processingcatalog configured to record operational data corresponding to one ormore discrete operational states of a point of service system. The oneor more discrete operational states may be selected from the groupconsisting of sample preparation, sample assaying and sample detection.Next, operational data of the point of service system is sequentiallyrecorded in the processing catalog.

In some cases, the operational data is recorded in real time. That is,the operational data may be recorded as a change or an update in anoperational state of the point of service system is detected or informedof.

In some cases, operational data is recorded in the sample processingcatalog prior to the point of service system performing a processingroutine corresponding to an operational state of the point of servicesystem. In other cases, operational data is recorded in the sampleprocessing catalog after the point of service system performs theprocessing routine. As an alternative, the operational data is recordedin the sample processing catalog while the point of service system isperforming the processing routine. In some cases, the log data isrecorded prior to, during and after completion of a transaction toprovide the most granular level of logging for every action across timeand space for the overall system level logging, or for the purpose ofsystem integrity and recovery.

The point of service system is configured to record the progress ofvarious processing routines of the point of service system and/orvarious components of the modules of the point of service system. Insome situations, the point of service system records in a processingcatalog when a processing routine has been completed by the point ofservice system.

A processing catalog may be provided by way of one or matrices stored onthe point of service system or another system associated with the pointof service system. In some situations, a point of service device (e.g.,the system 700 of FIG. 7) or module (e.g., the first module 701 of FIG.7) may include an operational matrix having discrete operational statesof the point of service device or module. The operational matrixincludes discrete states, namely State 1, State 2, State 3, and so on,of individual modules of a point of service system or components of amodule. The rows (if row matrix) or columns (if column matrix) of theoperational matrix are reserved for each module or component. Inaddition, each state may include one or more sub-states, and eachsub-state may include one or more sub-states. For instance, a modulehaving a first state, State 1, may have components performing variousfunctions. The states of various components have states designated byState mn, with ‘m’ designating the module and ‘n’ designating thecomponent of the module. In an example, for a first module of a point ofservice device, a first component may have a first state, State 11, anda second component may have a second state, State 12, and for a secondmodule of the point of service device, a first component may have afirst state, State 21, and a second component may have a second state,State 22. Each module may have any number of components (orsub-modules), such as at least one component (e.g., a singlecentrifuge), at least 10 components, or at least 100 components.

FIG. 42 shows an example of an operational matrix of a point of servicesystem, in accordance with an embodiment of the invention. Theoperational matrix may be for the point of service system or a module ofthe system or any component of the system or any module. The operationalmatrix includes a first column and a second the column, the first columnhaving numbers that correspond to the sequence number (“Sequence No.”)and the second column having strings that correspond to the operationalstate (e.g., “State 1”) of the system. Each operational state includesone or more routines, Routine n, wherein ‘n’ is an integer greater thanor equal to one. In the illustrated example, the first state (“State 1”)includes at least three routines, “Routine 1”, “Routine 2” and “Routine3.” In an embodiment, a routine includes one or more instructions thatindividually or in association with other routines bring the system ormodule in the system in-line with a particular state of the system.

A matrix may be located (or stored) on a physical storage medium of, orassociated with, a controller of a point of service device. The physicalstorage medium may be part of a database of the point of service device.The database may include one or more components selected from the groupconsisting of central processing unit (CPU), hard disk and memory (e.g.,flash memory). The database may be on-board the device and/or containedwithin the device. Alternatively, the data may be transmitted from adevice to an external device, and/or a cloud computing infrastructure.The matrix may be provided by way of one or more spreadsheets, datafiles having one or more rows and columns. Alternatively, the matrix canbe defined by one or more rows and one or more columns existing in amemory or other storage location of a controller or other system on orassociated with the point of service device.

FIG. 43 is an example of an operational matrix of a point of servicesystem and/or one or more modules of the point of service system. Theoperational matrix includes three processing states of the module,namely “Centrifuge sample,” “Perform cytometry on sample” and “Conductagglutination assay on sample.” Each processing state includes one ormore routines. For example, the first processing state (“Centrifugesample”) has six routines, as illustrated i.e., “Remove sample fromsample handling system”, “Provide sample in centrifugation tip”, and soon. The routines are listed in order of increasing time. That is, the“Remove sample from sample handling system” routine is performed beforethe “Provide sample in centrifugation tip” routine.

In some situations, operational data is provided in a one-dimensionalmatrix (i.e., column or row matrix). In other situations, operationaldata is provided in a two-dimensional matrix, with rows corresponding toroutines and columns corresponding to individual systems or systemmodules.

An operational matrix permits a point of service system to determinewhat processing routines have been conducted by the system at the mostgranular level of details in the system. This advantageously enables thesystem to recover from a fault condition in cases in which the systemrecords which processing routines were completed in a particular stateprior to a fault condition (e.g., power outage, system crash, modulecrash).

In some embodiments, a method for updating an operational log journal ofa point of service system comprises accessing an operational log journalof the point of service system, the operational log journal configuredto record operational data corresponding to one or more discreteoperational states of the point of service system. The operational logjournal may be accessed by the point of service system, a controller ofthe point of service system, or another system of the point of servicesystem or associated with the point of service system (collectively “thesystem”). The one or more discrete operational states include one ormore predetermined processing routines (e.g., centrifugation, PCR, oneor more assays). Next, the system generates one or more processingroutines to be performed by the point of service system. The processingroutines correspond to one or more operational states of the point ofservice system. The system then records data corresponding to the one ormore processing routines in the operational journal.

In some cases, the operational log journal may be part of an operatingsystem of the system. Alternatively, the operational log journal is asoftware or other computer-implemented application residing on thesystem or the cloud.

In some cases, the journal is implemented (or resides) on a hard disk ora flash drive that is not part of the hard disk. The journaling systemmay be separately powered by a battery in addition to the external powerto provide uninterrupted power supply to the journaling system in caseof system crash or disruptions of power from external or other sources.In other cases, the operational journal resides on a storage medium(hard disk, flash drive) of another system, such as a remote system.

In another embodiment, the log journaling is consulted when the systemboots up and resets the system. If the system has previously crashed orstopped abnormally, the system will use the log journal to bring allmodules, components and the system gracefully so the system can berelied upon. In some instances, the system may consult the log journalperiodically to monitor the status of each module, component,sub-component and so on and provide real-time recovery of any errors.

In another embodiment, the system may use log journal and onboardcameras to provide oversight over the entire system or a given module.In that case, the system may notice anomalies and missteps in real timeor near real-time and take corrective action. In another embodiment, thesystem may send these observations to an external device, such as acloud, and receive instructions from the external device on how toremedy any errors or missteps in the system.

In another embodiment, a method for processing a sample with the aid ofa point of service system comprises accessing an operational journal ofthe point of service system. The operational journal has operationaldata corresponding to one or more discrete operational states of thepoint of service system. The one or more discrete operational statesinclude one or more predetermined processing routines. The systemsequentially performs a processing routine from the one or morepredetermined processing routines, and removes, from the operationaljournal, data corresponding to a completed processing routine of anoperational state of the point of service system.

In an embodiment, the data corresponding to the completed processingroutine is removed from the operational journal before, during or aftersequentially performing the processing routine.

In some embodiments, a computer-assisted method for restoring anoperational state of a point of service system comprises accessing asample processing catalog following a fault condition of the point ofservice system; identifying a last-in-time operational state of thepoint of service system from the sample processing catalog; identifyinga last-in-time sample processing routine from said one or morepredetermined sample processing routines, the last-in-time sampleprocessing routine corresponding to the last-in-time operational stateof the point of service system; and performing a next-in-time processingroutine selected from the one or more predetermined sample processingroutines, the next-in-time processing routine following saidlast-in-time sample processing routine. The sample processing catalog isconfigured to record operational data corresponding to one or morediscrete operational states of the point of service system. In somecases, the operational data is recorded in the sample processing catalogfollowing the completion of a sample processing routine sequentiallyselected from one or more predetermined sample processing routines. Theone or more operational states of the point of service system areselected from the group consisting of sample preparation, sampleassaying and sample detection. In some cases, the fault condition isselected from the group consisting of a system crash, a power outage, ahardware fault, a software fault, and an operating system fault.

In other embodiments, a computer-assisted method for restoring anoperational state of a point of service system comprises accessing anoperational journal of the point of service system following a faultcondition of the point of service system. Next, one or more processingroutines corresponding to the operational data are sequentially replayedfrom the operational journal. The one or more processing routines arereplayed without the point of service system performing the one or moreprocessing routines. The system stops replaying the one or moreprocessing routines when a processing routine from the one or moreprocessing routines corresponds to an operational state of the point ofservice system prior to the fault condition. The system then restoresthe point of service system to the operational state prior to the faultcondition. In some cases, the operational journal has operational datacorresponding to one or more discrete operational states of the point ofservice system. The one or more discrete operational states include oneor more predetermined processing routines.

FIG. 44 shows a Plan matrix and a Routine matrix. The plan and routinematrices may be part of one or more operational matrices of a point ofservice system. The Plan matrix includes predetermined routines to beperformed by a point of service system or a module of the point ofservice system (“the system”). In some embodiments, the routines may bedynamic and may take into account, for example sample type, timing, orinformation relating to a system crash. The planned routines (“plans”)are sequentially listed, from top to bottom, in the order in which suchplans are to be performed by the system. The Routine matrix includesroutines (or plans) that have been performed by the system. As thesystem performs a particular routine, the system records the routine inthe routine matrix. Routines are recorded in the routine matrix in theorder in which they are performed. The routine at the bottom of the listis the routine that is performed last in time. In some situations, aroutine is marked as complete once one or more of the steps necessaryfor completing the routine have been completed by the system.

In an example, following a fault condition, the system accesses theroutine matrix to determine the routine performed last in time. Thesystem then begins processing with the plan (from the Plan matrix)selected following the routine last performed in time. In theillustrated example, the system begins processing by providing acentrifugation tip in the centrifuge. In some embodiments, faultrecovery may occur with information from an external device (e.g. thecloud).

In one embodiment, the system provides a pointer to indicate thelast-in-time processing routine to be completed prior to a faultcondition. FIG. 45A shows an operational matrix having processingstates. Each processing state has one or more routines in a Routinematrix. In the illustrated example, completed routines are shown inblack text and routines yet to be completed are shown in gray text. Theto-be-completed routines may be populated by reference to a Plan matrix,as described above. The horizontal arrow is a pointer marking theposition in the Routine matrix immediately following a last-in-timeroutine. Following a fault condition, the system begins processing atthe position indicated by the horizontal arrow. Here, the systemprovides a centrifugation tip in a centrifuge. In other cases, thesystem includes a pointer marking the position of a current andto-be-completed processing routine. In FIG. 45B, the horizontal arrow isa pointer marking the position of a processing routine (“Provide samplein centrifugation tip”) that has not been completed. The system may beperforming such processing routine between 0% but less than 100% tocompletion. Once complete, the horizontal arrow increments to the nextroutine (the arrow is incremented down along the Routine matrix).Following a fault condition, the system begins processing at theposition indicated by the horizontal arrow. As another alternative, thesystem includes a pointer marking the position of a processing routineto be completed immediately following a current processing routine. InFIG. 45C, the horizontal arrow is a point marking the position of aprocessing routine (“Provide centrifugation tip in centrifuge”) that isnext to be processed. In the illustrated example, the system is stillperforming the previous processing routine (“Provide sample incentrifugation tip”, as shown in gray). To-be-completed routines may bepopulated by reference to a Plan matrix, as described above.

In some embodiments, tracking processing routines may also includetracking precise locations of one or more components. Tracking aprocessing routine may include tracking each step or location involvedwith tracking the processing routine. For example, tracking a locationof a component may keep track of the exact distance (e.g., trackingevery mm, μm, nm, or less) that a component has moved. Even if acomponent has not yet reached its destination, the distance that it hastraveled on its journey may be tracked. Thus, even if an error occurs,the precise location of the component may be known and may be useful fordetermining the next steps. In another example, the amount of time anitem has been centrifuged may be tracked, even if the centrifuge processhas not yet been completed.

Components

A device may comprise one or more components. One or more of thesecomponents may be module components, which may be provided to a module.One or more of these components are not module components, and may beprovided to the device, but external to the module.

Examples of device components may include a fluid handling system, tips,vessels, microcard, assay units (which may be in the forms of tips orvessels), reagent units (which may be in the form of tips or vessels),dilution units (which may be in form of tips or vessels), wash units(which may in the form of tips or vessels), contamination reductionfeatures, lysing features, filtration, centrifuge, temperature control,detector, housing/support, controller, display/user interface, powersource, communication units, device tools, and/or device identifier.

One, two, or more of the device components may also be modulecomponents. In some embodiments, some components may be provided at boththe device level and module level and/or the device and module may bethe same. For example, a device may have its own power source, while amodule may also have its own power source.

FIG. 2 provides a high level illustration of a device 200. The devicemay have a housing 240. In some embodiments, one or more components ofthe device may be contained within the device housing. For example, thedevice may include one or more support structure 220, which may have oneor more module 230 a, 230 b. The device may also have a samplecollection unit 210. A device may have a communication unit 280 capableof permitting the device to communicate with one or more external device290. The device may also include a power unit 270. A device may have adisplay/user interface 260 which may be visible to an operator or userof the device. In some situations, the user interface 260 displays auser interface, such as graphical user interface (GUI), to a subject.The device may also have a controller 250 which may provide instructionsto one or more component of the device.

In some embodiments, the display unit on the device may be detachable.In some embodiments, the display unit may also have a CPU, memory,graphics processor, communication unit, rechargeable battery and otherperipherals to enable to operate it as a “tablet computer” or “slatecomputer” enabling it to communicate wirelessly to the device. In someembodiments, the detached display/tablet may be a shared source amongstall devices in one location or a group so one “tablet” can control,input and interact with 1, 2, 5, 10, 100, 1000 or more devices.

In some embodiments, the detached display may act as companion devicefor a healthcare professional to not only control the device, but alsoact as touch-enabled input device for capturing patient signatures,waivers and other authorizations and collaborating with other patientsand healthcare professionals.

The housing may surround (or enclose) one or more components of thedevice.

The sample collection unit may be in fluid communication with one ormore module. In some embodiments, the sample collection unit may beselectively in fluid communication with the one or more module. Forexample, the sample collection unit may be selectively brought intofluid communication with a module and/or brought out of fluidcommunication with the module. In some embodiments, the samplecollection unit may be fluidically isolated from the module. A fluidhandling system may assist with transporting a sample from a samplecollection unit to a module. The fluid handling system may transport thefluid while the sample collection unit remains fluidically orhydraulically isolated from the module. Alternatively, the fluidhandling system may permit the sample collection unit to be fluidicallyconnected to the module.

The communication unit may be capable of communicating with an externaldevice. Two-way communication may be provided between the communicationunit and the external device.

The power unit may be an internal power source or may be connected to anexternal power source.

Further descriptions of a diagnostic device and one or more devicecomponents may be discussed in greater detail elsewhere herein.

Fluid Handling System

A device may have a fluid handling system. As previously described, anydiscussion herein of fluid handling systems may apply to any samplinghandling system or vice versa. In some embodiments, a fluid handlingsystem may be contained within a device housing. The fluid handlingsystem or portions of the fluid handling system may be contained withina module housing. The fluid handling system may permit the collection,delivery, processing and/or transport of a fluid, dissolution of dryreagents, mixing of liquid and/or dry reagents with a liquid, as well ascollection, delivery, processing and/or transport of non-fluidiccomponents, samples, or materials. The fluid may be a sample, a reagent,diluent, wash, dye, or any other fluid that may be used by the device. Afluid handled by the fluid handling system may include a homogenousfluid, or fluid with particles or solid components therein. A fluidhandled by a fluid handling system may have at least a portion of fluidtherein. The fluid handling system may be capable of handlingdissolution of dry reagents, mixing of liquid and/or dry reagents in aliquid. “Fluids” can include, but not limited to, different liquids,emulsions, suspensions, etc. Different fluids may be handled usingdifferent fluid transfer devices (tips, capillaries, etc.). A fluidhandling system, including without limitation a pipette, may also beused to transport vessels around the device. A fluid handling system maybe capable of handling flowing material, including, but not limited to,a liquid or gaseous fluid, or any combination thereof. The fluidhandling system may dispense and/or aspirate the fluid. The fluidhandling system may dispense and/or aspirate a sample or other fluid,which may be a bodily fluid or any other type of fluid. The sample mayinclude one or more particulate or solid matter floating within a fluid.

In one example, the fluid handling system may use a pipette or similardevice. A fluid handling device may be part of the fluid handlingsystem, and may be a pipette, syringe, capillary, or any other device.The fluid handling device may have portion with an interior surface andan exterior surface and an open end. The fluid handling system may alsocontain one or more pipettes, each of which has multiple orificesthrough which venting and/or fluid flows may happen simultaneously. Insome instances, the portion with an interior surface and an exteriorsurface and open end may be a tip. The tip may or may not be removablefrom a pipette nozzle. The open end may collect a fluid. The fluid maybe dispensed through the same open end. Alternatively, the fluid may bedispensed through another end.

A collected fluid may be selectively contained within the fluid handlingdevice. The fluid may be dispensed from the fluid handling device whendesired. For example, a pipette may selectively aspirate a fluid. Thepipette may aspirate a selected amount of fluid. The pipette may becapable of actuating stirring mechanisms to mix the fluid within the tipor within a vessel. The pipette may incorporate tips or vessels creatingcontinuous flow loops for mixing, including of materials or reagentsthat are in non-liquid form. A pipette tip may also facilitate mixtureby metered delivery of multiple fluids simultaneously or in sequence,such as in 2-part substrate reactions. The fluid may be contained withina pipette tip, until it is desired to dispense through fluid from thepipette tip. In some embodiments, the entirety of the fluid within thefluid handling device may be dispensed. Alternatively, a portion of thefluid within the fluid handling device may be dispensed. A selectedamount of the fluid within the fluid handling device may be dispensed orretained in a tip.

A fluid handling device may include one or more fluid handling orificeand one or more tip. For example, the fluid handling device may be apipette with a pipette nozzle and a removable/separable pipette tip. Thetip may be connected to the fluid handling orifice. The tip may beremovable from the fluid handling orifice. Alternatively, the tip may beintegrally formed on the fluid handling orifice or may be permanentlyaffixed to the fluid handling orifice. When connected with the fluidhandling orifice, the tip may form a fluid-tight seal. In someembodiments, a fluid handling orifice if capable of accepting a singletip. Alternatively, the fluid handling orifice may be configured toaccept a plurality of tips simultaneously.

The fluid handling device may include one or more fluidically isolatedor hydraulically independent units. For example, the fluid handlingdevice may include one, two, or more pipette tips. The pipette tips maybe configured to accept and confine a fluid. The tips may be fluidicallyisolated from or hydraulically independent of one another. The fluidcontained within the tips may be fluidically isolated or hydraulicallyindependent from one another and other fluids within the device. Thefluidically isolated or hydraulically independent units may be movablerelative to other portions of the device and/or one another. Thefluidically isolated or hydraulically independent units may beindividually movable.

A fluid handling device may include one, two, three, four or more typesof mechanisms in order to dispense and/or aspirate a fluid. For example,the fluid handling device may include a positive displacement pipetteand/or an air displacement pipette. The fluid handling device mayinclude piezo-electric or acoustic dispensers and other types ofdispensers. The fluid handling device may include, one, two, three,four, five, six, seven, eight, nine, ten, or more positive displacementpipettes. The fluid handling device may be capable of metering(aspirating) very small droplets of fluid from pipette nozzles/tips. Thefluid handling device may include one or more, two or more, 4 or more, 8or more, 12 or more, 16 or more, 20 or more, 24 or more, 30 or more, 50or more, or 100 or more air displacement pipettes. In some embodiments,the same number of positive displacement pipettes and air displacementpipettes may be used. Alternatively, more air displacement pipettes maybe provided than positive displacement pipettes, or vice versa. In someembodiments, one or more positive displacement pipette can be integratedinto the “blade” style (or modular) pipetter format to save space andprovide additional custom configurations.

In some embodiments, a fluid handling apparatus, such as a pipette(e.g., a positive displacement pipette, air displacement pipette,piezo-electric pipette, acoustic pipette, or other types of pipettes orfluid handling apparatuses) described elsewhere herein, may have thecapability of picking up several different liquids with or withoutseparation by air “plugs.” A fluid handling apparatus may have thecapability of promoting/accelerating reaction with reagents attached tosurface (e.g., pipette tip surfaces) by reciprocating movement of theenclosed liquid, thus breaking down an unstirred layer. Thereciprocating movement may be driven pneumatically. The motion may beequivalent or comparable to orbital movement of microtiter places toaccelerate binding reactions in ELISA assays.

A fluid handling device may comprise one or more base or support. Thebase and/or support may support one or more pipette head. A pipette headmay comprise a pipette body and a pipette nozzle. The pipette nozzle maybe configured to interface with and/or connect to a removable tip. Thebase and/or support may connect the one or more pipette heads of thefluid handling device to one another. The base and/or support may holdand/or carry the weight of the pipette heads. The base and/or supportmay permit the pipette heads to be moved together. One or more pipettehead may extend from the base and/or support. In some embodiments, oneor more positive displacement pipette and one or more air displacementpipette may share a base or support.

Positive Displacement Pipette

FIG. 12 shows an exploded view of a positive displacement pipetteprovided in accordance with an embodiment of the invention. A positivedisplacement pipette may include a lower portion including a positivedisplacement pipette tip 1200, a nozzle 1202 and a slotted sleeve 1204.The positive displacement pipette may also include an inner portionincluding a collette 1212, collette sleeve 1214, collette spring 1216,and collette cap and hammer 1218. The positive displacement pipette mayinclude an upper portion including a screw helix 1220 with a hammer pin1222, a base 1228, and a DC gearmotor 1230.

A positive displacement pipette may permit the dispensing or aspirationof a fluid with a high degree of accuracy and precision. For example,using a positive displacement pipette, the amount of fluid dispensed oraspirated may be controlled to within about 1 mL, 500 microliters (μL,also “ul” herein), 300 μL, 200 μL, 150 μL, 100 μL, 50 μL, 30 μL, 10 μL,5 μL, 1 μL, 500 nL, 300 nL, 100 nL, 50 nL, 10 nL, 5 nL, 1 nL, 500 pL,100 pL, 10 pL, or 1 pL.

A positive displacement pipette may have a low coefficient of variation(CV). For example, the CV may be 10% or less, 8% or less, 5% or less, 3%or less, 2% or less, 1.5% or less, 1% or less, 0.7% or less, 0.5% orless, 0.3% or less, 0.1% or less, 0.05% or less, 0.01% or less, 0.005%or less, or 0.001% or less. In some cases, a positive displacementpipette having such a coefficient of variation may be configured tohandle sample (e.g., fluid) volumes less than or equal to 10 mL, 5 mL, 3mL, 2 mL, 1 mL, 0.7 mL, 0.5 mL, 0.4 mL, 0.3 mL, 250 μL, 200 μL, 175 μL,160 μL, 150 μL, 140 μL, 130 μL, 120 μL, 110 μL, 100 μL, 70 μL, 50 μL, 30μL, 20 μL, 10 μL, 7 μL, 5 μL, 3 μL, 1 μL, 500 nL, 300 nL, 100 nL, 50 nL,10 nL, 5 nL, 1 nL, 500 pL, 100 pL, 50 pL, 10 pL, 5 pL, 1 pL. In othercases, a positive displacement pipette having such a coefficient ofvariation is configured to handle sample volumes greater than 10 mL, 20mL, 30 mL, 40 mL, 50 mL, 100 mL, or higher.

A positive displacement pipette may cause the fluid to be dispensedand/or aspirated by trapping a fixed amount of the fluid, anddischarging it by altering the volume of the cavity in which the fluidis trapped. The positive displacement pipette may trap the fluid withoutalso trapping air. In another embodiment, a single pipette may becapable of trapping multiple quantities or types of liquid by separatingthe trapped liquids with “plugs” of air. The tip of the positivedisplacement pipette may include a plunger that may directly displacethe fluid. In some embodiments, the tip of the positive displacementpipette may function as a microsyringe, where the internal plunger maydirectly displace the liquid.

A positive displacement pipette may have a variety of formats. Forexample, the plunger may slide up and down based on various actuationmechanisms. The use of a screw helix 1220 with a hammer pin 1222 mayadvantageously permit a great degree of control on the volume aspiratedand/or dispensed. This may be very useful in situations where smallvolumes of fluid are handled. The screw helix may be mechanicallycoupled to a motor 1230. The motor may rotate, thereby causing the screwhelix to rotate. In some embodiments, the screw helix may be directlylinked to the motor so that the helix turns the same amount to that themotor turns. Alternatively, the screw helix may be indirectly coupled tothe motor so that the helix may turn some ratio relative to the amountthat the motor turns.

The hammer pin 1222 may be positioned through the screw helix 1220. Thehammer pin may have an orthogonal orientation in relation to the screwhelix. For example, if the screw helix is vertically aligned, the hammerpin may be horizontally aligned. The hammer pin may pass through thescrew helix at two points. In some embodiments, the screw helix andhammer pin may be positioned within a slotted sleeve 1204. An end of thehammer pin may fit within the slot of the sleeve. In some embodiments,the slotted sleeve may have two slots, and the hammer pin may have twoends. A first end of the hammer pin may be within a first slot of thesleeve, and a second end of the hammer may be within a second slot ofthe sleeve. The slots may prevent the hammer pin from rotating. Thus,when the screw helix is turned by a motor, the hammer pin may travel upand down along the slots.

As the hammer pin 1222 may optionally pass through a collet cap andhammer 1218. The collet cap may be directly or indirectly connected to acollet. The collet may be capable of passing into and through at least aportion of a pipette nozzle 1202. As the hammer pin may travel up anddown the slots, the collet may also travel up and down the slot. Thecollet pin may travel up and down the same amount that the hammer pintravels. Alternatively, the collet pin may travel some ratio of thedistance that the hammer pin travels. The collet pin may be directly orindirectly coupled to the hammer pin.

The collet preferably does not directly contact the fluid collected inand/or dispensed by a pipette tip. Alternatively the collet may contactthe fluid. The collet may contact a plunger that may preferably directlycontact the fluid collected in and/or dispensed by a pipette tip.Alternatively, the plunger may not directly contact the fluid. Theamount that the collet moves up and down may determine the amount offluid dispensed.

The use of a screw helix may provide a high degree of control of theamount of fluid dispensed and/or aspirated. A significant amount ofmotion rotating the screw may translate to a small amount of motion forthe hammer pin sliding up and down, and thus, the plunger within thepipette tip.

A positive displacement pipette may have a full aspiration position anda full dispense position. When the pipette is in a full aspirationposition, the collet may be at a top position. When the pipette is in afull dispense position, the collet may be at a bottom position. Thepipette may be capable of transitioning between a full aspiration and afull dispense position. The pipette may be capable of having anyposition between a full aspiration and full dispense position. Thepipette may have a partially aspirated and partially dispensed position.The pipette may stop at any in-between position smoothly in an analogmanner. Alternatively, the pipette may stop at particular in-betweenpositions with fixed increments in a digital manner. The pipette maymove from a dispense to aspirated position (e.g., have the colletassembly move upward toward the motor) in order to aspirate or draw afluid in. The pipette may move from an aspirated to a dispense position(e.g., have the collet assembly move downward away from the motor) inorder to dispense or eject some fluid out.

FIG. 13 shows an exterior side view and a side cross-section of apositive displacement tip in a top position (e.g., full aspirationposition). The pipette tip is not shown for clarity. A motor 1300 may becoupled to a helix 1310. The helix may be located beneath the motor. Thehelix may be located between a motor and a positive displacement tip. Acollet assembly 1320 may be located within the helix. The helix may wraparound, or surround, the collet assembly.

A plunger spring 1330 may be provided between the collet assembly 1320and the helix 1310. The collet assembly may have a shelf or protrudingportion, upon which one end of the plunger spring may be supported, orrest. The pipette nozzle 1340 may have another shelf or protrudingportion upon which one end of the plunger spring may be supported orrest. The plunger spring may be located between a pipette nozzle, and atop portion of a collet.

When a positive displacement pipette is in its full aspiration position,the plunger spring may be in an extended state. The plunger spring maykeep a collet assembly at an upper position, when the pipette is in anaspirated position.

FIG. 14 shows an exterior side view and a side cross-section of apositive displacement tip in a bottom position (e.g., full dispenseposition). A motor 1400 may be coupled to a helix 1410. The helix may belocated beneath the motor. The helix may be located between a motor anda positive displacement tip. A collet assembly 1420 may be locatedwithin the helix or at least partially beneath the helix. The helix maywrap around, or surround, the collet assembly.

A plunger spring 1430 may be provided at least partially between thecollet assembly 1420 and the helix 1410. The collet assembly may have ashelf or protruding portion, upon which one end of the plunger springmay be supported, or rest. The pipette nozzle 1440 may have anothershelf or protruding portion upon which one end of the plunger spring maybe supported or rest. The plunger spring may be located between apipette nozzle, and a top portion of a collet. The plunger spring maysurround at least a portion of the collet assembly.

When a positive displacement pipette is in its full dispense position,the plunger spring may be in a compressed state. The collet assembly maybe driven downward toward the tip, thereby compressing the spring. Thepipette may have two (or more) plungers and/or collets that enableaspiration/dispensing of two materials and subsequent mixing; forexample, processing of an epoxy, which is a copolymer that is formedfrom two different chemicals; the mixing and metering can be finelycontrolled with respect to volumes and times.

A positive displacement tip plunger 1450 may be connected to the colletassembly 1420. The plunger may be located beneath the collet assembly.The plunger may be located between the collet assembly and the tip. Thepositive displacement tip plunger may include an elongated portion thatmay be capable of extending at least partially through the pipette tip.In some embodiments, the elongated portion may be long enough to extendcompletely through the pipette tip when in a full dispense position. Insome embodiments, when in full dispense position, the elongated portionof the plunger may extend beyond the pipette tip. The end of the plungermay or may not directly contact a fluid aspirated and/or dispensed bythe positive displacement pipette. In some embodiments, the plunger mayhave a protruding portion or shelf that may rest upon the colletassembly. The plunger may move up and down the same amount that a colletassembly moves up and down.

The pipette tip may have any configuration of tips as describedelsewhere herein. For example, the pipette tip may have a positivedisplacement tip as illustrated by FIG. 14 or FIG. 27. The positivedisplacement tip may be configured to confine and accept any volume offluid, including those described elsewhere herein.

Referring now to FIG. 91, one embodiment of an engagement mechanism fora positive displacement (PD) tip will now be described. As seen in FIG.91, the ‘collet driver’ 1460 can be magnetically or mechanicallyconnected to any mechanism that can drive it relative to a stationary‘housing’ 1462. This embodiment may include a compression spring 1464, acollet sleeve 1466, and the collet 1468. The motion of the collet driver1460 causes the same effect in the rest of the system as the motion ofthe ‘hammer pin’ in the ‘helix’ in the other embodiments of the PD tips.By having this setup with the compression spring 1464 inside the colletsleeve 1466, the drive actuator does not have to be in-line with the PDassembly, and the PD mechanism is independent of actuation method. Theentire assembly may be part of the pipette mechanism.

Air Displacement Pipette

FIG. 15 shows an exterior view of an air displacement pipette providedin accordance with an embodiment of the invention. An air displacementpipette may include a pipette tip 1500 and an external removal mechanism1510 for removing the pipette tip from a pipette nozzle 1520.

An air displacement pipette may permit the dispensing or aspiration of afluid with a high degree of accuracy and precision. For example, usingan air displacement pipette, the amount of fluid dispensed or aspiratedmay be controlled to within about 3 mL, 2 mL, 1.5 mL, 1 mL, 750 μL, 500μL, 400 μL, 300 μL, 200 μL, 150 μL, 100 μL, 50 μL, 30 μL, 10 μL, 5 μL, 1μL, 500 nL, 300 nL, 100 nL, 50 nL, 10 nL, or 1 nL. In some embodiments,a positive displacement pipette may have a higher degree of accuracyand/or precision than an air displacement pipette.

In some embodiments, one or more pipettes, such as one or more of an airdisplacement pipette, positive displacement pipette and suction-typepipette, may have a low coefficient of variation (CV). For example, theCV may be 15% or less, 12% or less, 10% or less, 8% or less, 5% or less,3% or less, 2% or less, 1.5% or less, 1% or less, 0.7% or less, 0.5% orless, 0.3% or less, or 0.1% or less. In some cases, a pipette (e.g.,positive displacement pipette, air displacement pipette, or suction-typepipette) having such a coefficient of variation may be configured tohandle sample (e.g., fluid) volumes less than or equal to 10 mL, 5 mL, 3mL, 2 mL, 1 mL, 0.7 mL, 0.5 mL, 0.4 mL, 0.3 mL, 250 μL, 200 μL, 175 μL,160 μL, 150 μL, 140 μL, 130 μL, 120 μL, 110 μL, 100 μL, 70 μL, 50 μL, 30μL, 20 μL, 10 μL, 7 μL, 5 μL, 3 μL, 1 μL, 500 nL, 300 nL, 100 nL, 50 nL,10 nL, 5 nL, 1 nL, 500 pL, 100 pL, 50 pL, 10 pL, 5 pL, 1 pL. In othercases, a pipette (e.g., positive displacement pipette, air displacementpipette, or suction-type pipette) having such a coefficient of variationis configured to handle sample volumes greater than 10 mL, 20 mL, 30 mL,40 mL, 50 mL, 100 mL, or higher. Various types and combinations ofpipettes provided herein (e.g., positive displacement pipette, airdisplacement pipette, or suction-type pipette) are configured to havesuch a coefficient of variation while handling the sample volumes setforth herein.

An air displacement pipette may have a low coefficient of variation(CV). For example, the CV may be 10% or less, 8% or less, 5% or less, 3%or less, 2% or less, 1.5% or less, 1% or less, 0.7% or less, 0.5% orless, 0.3% or less, 0.1% or less, 0.05% or less, 0.01% or less, 0.005%or less, or 0.001% or less. In some cases, an air displacement pipettehaving such a coefficient of variation may be configured to handlesample (e.g., fluid) volumes less than or equal to 10 mL, 5 mL, 3 mL, 2mL, 1 mL, 0.7 mL, 0.5 mL, 0.4 mL, 0.3 mL, 250 μL, 200 μL, 175 μL, 160μL, 150 μL, 140 μL, 130 μL, 120 μL, 110 μL, 100 μL, 70 μL, 50 μL, 30 μL,20 μL, 10 μL, 7 μL, 5 μL, 3 μL, 1 μL, 500 nL, 300 nL, 100 nL, 50 nL, 10nL, 5 nL, 1 nL, 500 pL, 100 pL, 50 pL, 10 pL, 5 pL, 1 pL. In othercases, an air displacement pipette having such a coefficient ofvariation is configured to handle sample volumes greater than 10 mL, 20mL, 30 mL, 40 mL, 50 mL, 100 mL, or higher.

An air displacement pipette may cause the fluid to be dispensed and/oraspirated by generating a vacuum by the travel of a plunger within anair-tight sleeve. As the plunger moves upward, a vacuum is created inthe space left vacant by the plunger. Air from the tip rises to fill thespace left vacant. The tip air is then replaced by the fluid, which maybe drawn into the tip and available for transport and dispensingelsewhere. In some embodiments, air displacement pipettes may be subjectto the changing environment, such as temperature. In some embodiments,the environment may be controlled in order to provide improved accuracy.

The air displacement pipette may have a variety of formats. For example,the air displacement pipette may be adjustable or fixed. The tips may beconical or cylindrical. The pipettes may be standard or locking. Thepipettes may be electronically or automatically controlled, or may bemanual. The pipettes may be single channeled or multi-channeled.

FIG. 16 shows a cross-sectional view of air displacement pipette. Theair displacement pipette may include a pipette tip 1600 and an externalremoval mechanism 1610 for removing the pipette tip from a pipettenozzle 1620. The removal mechanism may be positioned to contact an endof the pipette tip. The removal mechanism may be positioned above thepipette tip at the end opposing an end of the pipette tip that dispensesand/or aspirates a fluid. The pipette tip may have a shelf or protrudingportion upon which the removal mechanism may rest.

The pipette tip may have any format of any tip as described elsewhereherein. For example, the tip may be a nucleic acid tip, centrifugationextraction tip, bulk handling tip, color tip, blood tip, minitip,microtip, nanotip, fentotip, picotip, and the like, or may have featuresor characteristics of any tips described in FIGS. 24 to 34.

FIG. 17 shows a close-up of an interface between a pipette tip 1700 anda nozzle 1720. A removal mechanism 1710 may be positioned to contact thepipette tip.

A pipette nozzle may have a protruding portion 1730 or a shelf that maycontact a removal mechanism. The nozzle shelf may prevent the removalmechanism from traveling too far upwards. The nozzle shelf may provide adesired position for the removal mechanism.

A pipette nozzle may also have one or more sealing element 1740. Thesealing elements may be one or more O-rings or other similar materialsknown in the art. The sealing elements may contact a pipette tip whenthe pipette tip is attached to the nozzle. The sealing element maypermit a fluid-tight seal to be formed between the pipette tip and thenozzle. The sealing element may keep the pipette tip attached to thenozzle in the absence of an outside force. The pipette tip may befriction-fit to the pipette nozzle.

An interior channel 1750 or chamber may be provided within the pipettenozzle. The pipette tip may have an interior surface 1760 and interiorregion 1770. The interior channel of the pipette nozzle may be in fluidcommunication with the interior region of the pipette tip. A plunger maytravel through the channel of the pipette nozzle and/or the interiorregion of the pipette tip. The plunger may permit the aspiration ordispensing of a fluid from the pipette tip. The plunger may or may notdirectly contact the fluid. In some embodiments, air may be providedbetween the plunger and the fluid.

FIG. 18 shows an example of an actuation of a removal mechanism 1810.The removal mechanism may cause a pipette tip 1800 to be removed from anozzle 1820. The removal mechanism may be provided external to thepipette tip and nozzle. The removal mechanism may be moved downward, inorder to push the pipette tip off the nozzle. Alternatively, the pipettenozzle may be moved upward, causing the pipette tip to be caught on theremoval mechanism and pushed off. The removal mechanism may moverelative to the pipette nozzle.

The removal mechanism may contact a pipette tip at the top of thepipette tip. The removal mechanism may contact the pipette tip on a sideof the pipette tip. The removal mechanism may go partially or completelyaround the pipette tip.

FIG. 19A shows a plurality of pipettes with an external removalmechanism. For example, eight pipette heads may be provided. In otherembodiments of the invention, any other number of pipette heads,including those described elsewhere herein, may be used.

FIG. 19B shows a side view of a pipette head. The pipette head mayinclude a pipette tip 1900. The pipette tip may be removable coupled toa pipette nozzle 1920. An external removal mechanism 1910 may beprovided. The external removal may be in contact with the pipette tip ormay come into contact with the pipette tip. The pipette nozzle may becoupled to a support 1930 of the pipette. The pipette support may becoupled to a motor 1940 or other actuation mechanism.

FIGS. 20A to C show cross-sectional views of an air displacementpipette. The air displacement pipette may include a pipette tip 2000 andan external removal mechanism 2010 for removing the pipette tip from apipette nozzle 2020. The removal mechanism may be positioned to contactan end of the pipette tip. The removal mechanism may be positioned abovethe pipette tip at the end opposing an end of the pipette tip thatdispenses and/or aspirates a fluid. The pipette tip may have a shelf orprotruding portion upon which the removal mechanism may rest.

The removal mechanism 2010 may travel up and down to remove a pipettetip from a nozzle. The removal mechanism may be coupled to an actuationmechanism that may permit the removal mechanism to travel up and down.In some embodiments, the removal mechanism may be directly coupled tothe actuation mechanism. Alternatively, the removal mechanism may beindirectly coupled to the actuation mechanism. One or more switch may beprovided between a removal mechanism and an actuation mechanism that maydetermine whether the removal mechanism responds to the actuationmechanism. The switch may be a solenoid or other mechanism.

The air displacement pipette may also include an internal plunger 2030.The plunger may travel through an interior portion of a pipette nozzle.The plunger may be coupled to an actuation mechanism that may permit theplunger to travel up and down. In some embodiments, the plunger may bedirectly coupled to the actuation mechanism. Alternatively, the plungermay be indirectly coupled to the actuation mechanism. One or more switchmay be provided between a plunger and an actuation mechanism that maydetermine whether the plunger responds to the actuation mechanism. Theswitch may be a solenoid or other mechanism.

FIG. 20A shows a plunger in a down position, as well as a removalmechanism in a down position, thereby pushing a tip down relative to thepipette nozzle.

FIG. 20B shows a plunger in an intermediate position, as well as aremoval mechanism in an up position, thereby permitting a tip to beattached to the pipette nozzle.

FIG. 20C shows a plunger in an up position, as well as a removalmechanism in an up position, thereby permitting a tip to be attached tothe pipette nozzle.

FIG. 21 shows a plurality of pipettes with removal mechanisms. Forexample, eight pipette heads may be provided. In other embodiments ofthe invention, any other number of pipette heads, including thosedescribed elsewhere herein, may be used.

A support structure 2100 for the pipettes may be provided. One or morepipette sleeve 2110 may be provided through which a plunger may extend.A spring 2120 may be provided in accordance with an embodiment of theinvention. The spring may be compressed when the plunger is moved down.The spring may be extended when the plunger is an up position.

One or more switching mechanisms, such as solenoids 2130 may beprovided. An actuation mechanism, such as a motor 2140 may be providedfor the plurality of pipettes. The actuation mechanism may be coupled tothe plungers and/or removal mechanisms of the pipettes. In someembodiments, the actuation mechanisms may be directly coupled to theplungers and/or removal mechanisms. Alternatively, the actuationmechanisms may be indirectly connected to the plungers and/or removalmechanisms. In some embodiments, one or more switches may be providedbetween the actuation mechanism and the plunger and/or removalmechanism. The switch may determine whether the plunger and/or removalmechanism responds to the actuation mechanism. In some embodiments, theswitches may be solenoids.

In some embodiments, a single actuation mechanism may be used to controleach of the pipettes pistons for the multi-head pipette. Switches may beprovided for each of the pipette pistons so that actuation may beindividually controllable for each of the pipette pistons. In someembodiments, the pipette piston can dynamically change its volume,thereby optimizing performance for the required sample volumes to beaspirated/dispensed; for example, the piston can be a tube within a tubethat is expandable to dynamically control volume. In some embodiments,switches may be provided for groups of pipette pistons so that theactuation may be individually controllable between each of the groups ofpipette pistons. A single actuation mechanism may be used to controleach of the pipette pistons. In some embodiments, single actuationmechanisms may be used to control groups of pipette pistons.Alternatively, each pipette piston may be connected to its ownindividual actuation mechanism. Thus, one, two, three, four or moreactuation mechanisms, (such as motors) may be provided for a pipettepiston.

FIG. 22 shows an example of a multi-head pipette in accordance with anembodiment of the invention. The individual pipette heads on themulti-head pipette may be individually actuatable or may haveindividually actuatable components. For example, a removal mechanism2200 for one of the pipette heads may be in an up position, while theother removal mechanisms 2210 may be in a down position. A switch, suchas a solenoid 2220, may be disengaged for that one pipette head, whilethe switches may be engaged for the other pipette head. Thus, when anactuation mechanism, such as a motor 2230, is engaged to move theremoval mechanisms downward to remove pipette tips from pipette nozzles,the one disengaged switch may cause that one removal mechanism to notmove downward with the others. The disengaged removal mechanism mayremain in its place. This may cause the pipette tip to remain on thedisengaged pipette, while pipette tips are removed from other pipettes.

In another example, a plunger 2250 for one of the pipette heads may bein an up position, while the other plungers 2260 may be in a downposition. A switch, such as a solenoid, may be disengaged for that onepipette head, while the switches may be engaged for the other pipettehead. Thus, when an actuation mechanism, such as a motor, is engaged tomove the plungers downward to dispense fluid or to remove pipette tipsfrom pipette nozzles, the one disengaged switch may cause that oneplunger to not move downward with the others. The disengaged plunger mayremain in its place. This may cause the pipette tip to remain on thedisengaged pipette, while pipette tips are removed from other pipettes,or may prevent fluid from being dispensed from the disengaged pipettewhile fluid is dispensed at other pipettes.

In some embodiments, a disengaged switch may prevent a pipette tip frombeing removed, or fluid from being dispensed. In some embodiments, adisengaged switch may prevent a pipette tip from being picked up. Forexample, the engaged switches may cause pipette heads to actuatedownward to pick up a pipette tip, while pipette heads coupled withdisengaged switches remain in a retracted position. In another example,engaged switches may cause one or more mechanism that picks up a pipettehead to actuate to pick up the head while disengaged switches preventone or more pipette tip pick-up mechanism from operating.

In some additional embodiments, a disengaged switch may prevent apipette tip from aspirating a fluid. For example, engaged switches maycause an internal plunger or other mechanism to move upwards to aspiratea fluid. A disengaged switch may cause a plunger to remain in its place.Thus, aspiration of fluids in multi-head pipettes may be individuallycontrolled while using one or more actuation mechanism.

A removal mechanism may be provided external to a pipette nozzle, orinternal to the pipette nozzle. Any description herein of any type ofremoval mechanism may also refer to other types of removal mechanisms.For example, descriptions of individually actuatable external removalmechanisms may also apply to internal removal mechanisms that may have aplunger form or other form that may be provided within a nozzle.

An actuation mechanism may be configured to actuate components in aplurality of pipettes. For example, an actuation mechanism may beconfigured to actuate removal mechanisms. An actuation mechanism may becable of actuating both a first removal mechanism and a second removalmechanism. A first solenoid may be operatively provided between theactuation mechanism and the first removal mechanism. A second solenoidmay be operatively provided between the actuation mechanism and thesecond removal mechanism. The first solenoid may be engaged ordisengaged to determine whether actuation by the actuation mechanism maycause movement of the removal mechanism. The second solenoid may beengaged or disengaged to determine whether actuation by the actuationmechanism may cause movement of the removal mechanism. The first andsecond solenoids may be engaged or disengaged independent of oneanother. Each of the solenoids for individual pipettes or groups ofpipettes controlled by an actuation mechanism may be engaged ordisengaged in response to one or more signals received from acontroller.

In some embodiments, the actuation mechanism may be located on the topof a pipette. The actuation mechanism may be located on a supportstructure at an end opposing the pipette tips. The actuation mechanismmay be located on a support structure at an end opposing the pipettenozzles. The actuation mechanism may comprise one or more shaft that maybe oriented parallel to one or more pipette tip. The actuation mechanismmay have an axis of rotation that may be parallel to an axis extendingalong the height of one or more pipette tip.

FIG. 23 shows an example of a multi-head pipette 2300 provided inaccordance with another embodiment of the invention. An actuationmechanism 2310 may be located on any portion of a pipette. For example,the actuation mechanism may be located on a side of the supportstructure. Alternatively it may be located on a top or bottom portion ofa support structure. The actuation mechanism may be located to a side ofthe support structure opposing the pipette nozzles 2320. The actuationmechanism may comprise one or more shaft 2330 that may be orientedperpendicular to one or more pipette tip 2340. The actuation mechanismmay have an axis of rotation that may be perpendicular to an axisextending along the height of one or more pipette tip. For example, apipette tip may have a vertical orientation, while an actuationmechanism may have a shaft or axis of rotation having a horizontalorientation. Alternatively, the actuation mechanism shaft or axis may beat any angle relative to the one or more pipette tip.

One or more pipette head or pipette support 2350 may have a bentconfiguration. For example, a pipette support may have a horizontalcomponent 2350 a that meets a vertical component 2350 b. In someembodiments, fluid may only be provided to a vertical component of thepipette. Alternatively, fluid may or may not flow to a horizontalcomponent of the pipette. A pipette may have a single piston or plungerthat can be linked to two or more nozzles or tips and a valve or switchcan be used to enable aspiration/dispensing through one or more of thenozzles or tips.

One or more switches 2360 may be provided. The switches may beindividually controllable. Examples of switches and controls asdescribed elsewhere herein may apply. The actuation mechanism may becapable of driving one or more pipette actuation component, such aspipette tip remover, one or more pipette tip mounter, one or more fluiddispensing mechanism, and/or one or more fluid aspirating mechanism. Theswitches may determine whether one or more of the pipette actuationcomponents are moved or not.

Having a side mounted actuation mechanism may reduce one or moredimensions of the multi-head pipette. For example, a side mountedactuation mechanism may reduce the vertical dimension of the multi-headpipette while maintaining the same barrel volume, and hence pipettecapacity. Depending on the desired placement of the pipette within thedevice and/or module or other constraints in the device and/or module, atop mounted, side mounted, or bottom mounted actuation mechanism may beselected.

Having a single actuation mechanism that causes the actuation of all thepipette actuation components may also reduce the dimensions for themulti-head pipette. A single actuation mechanism may control a pluralityof the pipette actuation components. In some embodiments, one or moreactuation mechanisms may be provided to control a plurality of pipetteactuation components.

FIG. 46 shows an example of a fluid handling apparatus in a collapsedposition, provided in accordance with another embodiment of theinvention. The fluid handling apparatus may include one or more tips4610, 4612, 4614. In some embodiments, a plurality of tip types may beprovided. For example, a positive displacement tip 4610 may be provided,an air displacement nozzle tip 4612, and an air displacement mini-nozzletip 4614 may be provided. A base 4620 may be provided, supporting one ormore pipette head. A positive displacement motor 4630 may be coupled toa positive displacement pipette head 4635.

A fluid handling apparatus may include a manifold 4640. The manifold mayinclude one or more vent ports 4642. A vent port may be fluidicallyconnected to the fluid path of a pipette head. In some embodiments, eachpipette head may have a vent port. In some instances, each airdisplacement pipette head may have a vent port. A tubing 4644 may beconnected to the manifold. The tubing may optionally connect themanifold to a positive or negative pressure source, ambient air, or areversible positive/negative pressure source.

A thermal spreader 4650 may be provided for a fluid handling apparatus.The thermal spreader may provide isothermal control. In someembodiments, the thermal spreader may be in thermal communication with aplurality of pipette heads. The thermal spreader may assist withequalizing temperature of the plurality of pipette heads.

A fluid handling apparatus may have one or more support portion. In someembodiments, the support portion may include an upper clamshell 4660 anda lower clamshell 4665.

FIG. 46A shows a collapsed fluid handling apparatus as previouslydescribed, in a fully retracted position. FIG. 46B shows a collapsedfluid handling apparatus, in a full z-drop position. In a full z-dropposition, an entire lower clamshell 4665 may be lowered relative to theupper clamshell 4660. The lower clamshell may support the pipette headsand nozzles. The pipette heads and nozzles may move with the lowerclamshell. The lower clamshell may include a front portion 4667 whichsupports the pipette heads, and a rear portion 4668 which supports anactuation mechanism and switching mechanisms.

FIG. 47 shows an example of a fluid handling apparatus in an extendedposition in accordance with an embodiment of the invention. The fluidhandling apparatus may include one or more tips 4710, 4712, 4714. Apositive displacement tip 4710 may be provided, an air displacementnozzle tip 4712, and an air displacement mini-nozzle tip 4714 may beprovided. The fluid handling apparatus may also include one or morenozzles 4720, 4722, 4724. A positive displacement nozzle 4720, an airdisplacement nozzle 4722, and an air displacement mini-nozzle 4724 maybe provided. The nozzles may interface with their respective tips. Insome instances, the nozzles may connect to their respective tips viapress-fit or any other interface mechanism. One or more tip ejector4732, 4734 may be provided. For example, a regular tip ejector 4732 maybe provided for removing an air displacement tip 4712. One or moremini-ejector 4734 may be provided for removing an air displacementmini-tip 4714. The ejectors may form collars. The ejectors may bedesigned to push the tips off. The ejectors may be located beneath thenozzles.

The fluid handling apparatus may be in a full z-drop position with alower clamshell 4765 lowered relative to an upper clamshell 4760.Furthermore, a z-clutch-bar 4770 may be provided which may engage any orall of the pipettes for individualized and/or combined nozzle drop (i.e.nozzle extension). FIG. 47 shows an example where all nozzles aredropped. However, the nozzles may be individually selectable todetermine which nozzles to drop. The nozzles may drop in response to asingle actuation mechanism, such as a motor. A switching mechanism maydetermine which pipettes are engaged with the bar. The clutch bar 4770illustrated shows the nozzles in a dropped position, with the clutch barlowered. A z-motor encoder 4780 may be provided. The encoder may permitthe tracking of the location of the motor movement.

An x-axis slider 4790 may be provided in accordance with someembodiments. The x-axis slider may permit the fluid handling apparatusto move laterally. In some embodiments, the fluid handling apparatus mayslide along a track.

FIG. 48 shows a front view of a fluid handling apparatus. A protectorplate 4810 may be provided in some embodiments. The protector plate mayprotect portions of the pipette head. The protector plate may protect afluid path of the pipette head. In one example, the protector plate maybe provided for rigid tubing, connecting pipette cavities to nozzles.The protector plate may be connected to a thermal spreader or may beintegrally incorporated with a thermal spreader.

As previously described, multiple types of pipettes and/or tips may beprovided. One or more positive displacement pipette and/or one or moreair displacement pipettes may be used. In some instances, the protectorplate may only cover the air displacement pipettes. Alternatively, theprotector place may cover the positive displacement pipette only, or maycover both.

FIG. 49 shows a side view of a fluid handling apparatus. A fluidhandling apparatus may include a pipette head, which may include anozzle head 4902, which may be configured to connect to a tip 4904. Thetip may be removably connected to the pipette nozzle.

One or more pipette nozzle may be supported by a nozzle-drop shaft 4920.A z-motor 4922 may be provided, which when actuated, may cause one ormore nozzle to drop (e.g., extend). One or more solenoid 4924, or otherswitching mechanism may be provided to selectively connect the z-motorwith the nozzle-drop shaft. When the solenoid is in an “on” position,actuation of the z-motor may cause the nozzle-drop shaft to be loweredor raised. When the solenoid is in an “off” position, actuation of thez-motor does not cause movement of the nozzle-drop shaft.

Tubing 4910 may be provided through the pipette head, and terminating atthe pipette nozzle. The tubing may have a portion with rigid innertubing 4910 a, and rigid outer tubing 4910 b. In some instances, therigid inner tubing may be movable while the rigid outer tubing isstationary. In other embodiments, the rigid inner tubing may be movableor stationary, and the rigid outer tubing may be movable or stationary.In some embodiments, the inner tubing may be movable relative to theouter tubing. The overall length of the tubing may be variable.

A plunger 4930 may be provided within the fluid handling apparatus. Theplunger may provide metering within a pipette cavity. An extension ofthe pipette cavity 4935 may be provided. In some instances, theextension of the pipette cavity may be in fluid communication with thetubing 4910. Alternatively, the tubing and the pipette cavity are not influid communication. In some embodiments, the pipette cavity and thetubing are parallel to one another. In other embodiments, the pipettecavity and the tubing are substantially non-parallel to one another.They may be substantially perpendicular to one another. The tubing mayhave a substantially vertical orientation while the pipette cavity mayhave a substantially horizontal orientation, or vice versa. In someembodiments, a pipette and tip may act in a push/pull fashion, such asin a multi-lumen tubing arrangement, to aspirate and dispensesimultaneously or sequentially.

One or more valves 4937 may be provided for controlling vent port accessto the pipettes. In some instances, a valve may correspond to anassociated pipette. A valve may determine whether a vent port is open orclosed. A valve may determine the degree to which a vent port is open.The vent port may be in communication with a pressure source, such as apositive or negative pressure source. The vent port may be incommunication with ambient air. The vent port may provide access to atubing 4910 from a manifold.

A clutch-bar 4940 for individual metering may be provided. The clutchbar may be connected to a motor that may be used to drive the meteringof the fluid. A guide shaft 4942 may optionally be provided. One or moresolenoid 4945 or other switching mechanism may be provided incommunication with the clutch-bar. The solenoid or other switchingmechanism may be provided to selectively connect the motor with theplunger 4930. When the solenoid is in an “on” position, actuation of themetering motor may cause the plunger to be engaged and move to dispenseand/or aspirate a fluid. When the solenoid is in an “off” position,actuation of the metering motor does not cause movement of the plunger.A plurality of plungers may be provided, each being associated with itsrespective solenoid, which may selectively be in an on or off position.Thus, when a motor is actuated, only the plungers associated with “on”solenoids may respond.

FIG. 50 shows another side view of a fluid handling apparatus. The viewincludes a view of the motor 5010 used for metering. The motor may beused for metering fluid within the air displacement pipettes. An encoder5020 for the motor is also illustrated. The encoder may permit thetracking of the motor movement. This ensures that the plunger positionis known at all times.

FIG. 51 shows a rear perspective view of a fluid handling apparatus. Thefluid handling apparatus may include a pump 5110. The pump may be influid communication with a pipette cavity. In some instances, the pumpmay be brought into or out of fluid communication with the pipettecavity. The pump may be in fluid communication with a manifold, and/orvent port. The pump may pump (or effect the movement of) liquid or air.

The pump may provide positive pressure and/or negative pressure. Thepump may be a reversible pump that may be capable of providing bothpositive and negative pressure. The pump may be actuated in pipettescontaining pistons to permit the pipette to aspirate or dispense anyvolume of liquid, without limitation by the positive displacement that agiven piston size is capable of generating. This factor, in combinationwith large volume tips, could permit a small pipette to aspirate ordispense large volumes of liquid for certain applications. The pump maypermit the pipette to function without motor or piston, while stillproviding fine control through pulse-width modulation.

A fluid handling apparatus may also include an accumulator 5120 with oneor more valves that may connect to a pressure source or ambientconditions. The accumulator may optionally connect to the reversiblepump, which may provide positive or negative pressure.

A multi-headed fluid handling apparatus, such as a multi-headed pipettemay be capable of picking up multiple tips/vessels on several pipettenozzles at the same time. For example, multiple pipette nozzles mayextend to pick up multiple tips/vessels. The multiple pipette nozzlesmay be individually controllable to determine which tips/vessels arepicked up. Multiple tips/vessels may be picked up simultaneously. Insome instances, all pipette nozzles may pick up pipette tips/vesselssubstantially simultaneously.

In some embodiments, pipettes do not include plungers. A sample (e.g.,fluid) may be moved in or with the aid of the pipette using positiveand/or negative pressure. In some situations, positive or negativepressure is provided with the aid of a gas or vacuum, respectively.Vacuum may be provided using a vacuum system having one or more vacuumpumps. Positive pressure may be provided with the aid of pressurizedair. Air may be pressurized using a compressor.

Dimensions/Ranges

One or more dimensions (e.g., length, width, or height) of a pipette maybe less than or equal to about 1 mm, 5 mm, 10 mm, 15 mm, 20 mm, 25 mm,30 mm, 35 mm, 40 mm, 45 mm, 50 mm, 55 mm, 60 mm, 70 mm, 80 mm, 90 mm,100 mm, 112 mm, 12 cm, 15 cm, 20 cm, 25 cm, 30 cm, or any otherdimension described elsewhere herein. One or more dimensions may be thesame, or may vary. For example, the height of a pipette may not exceed 1mm, 1 cm, 2 cm, 3 cm, 4 cm, 5 cm, 5.5 cm, 6 cm, 6.5 cm, 7 cm, 8 cm, 9cm, 10 cm, 11 cm, 12 cm, 13 cm, 15 cm, 17 cm, 20 cm, 25 cm, or 30 cm.

In some embodiments, a pipette may have a total volume of 1 cm³ or less,5 cm³ or less, 8 cm³ or less, 10 cm³ or less, 15 cm³ or less, 20 cm³ orless, 25 cm³ or less, 30 cm³ or less, 35 cm³ or less, 40 cm³ or less, 50cm³ or less, 60 cm³ or less, 70 cm³ or less, 80 cm³ or less, 90 cm³ orless, 100 cm³ or less, 120 cm³ or less, 150 cm³ or less, 200 cm³ orless, 250 cm³ or less, 300 cm³ or less, or 500 cm³ or less.

The pipette may have one or more pipette head. In some embodiments, anindividual pipette head of the pipette may be able to dispense anyvolume of fluid. For example, the individual pipette head may be capableof dispensing and/or aspirating fluids of no more than and/or equal toabout 10 mL, 5 mL, 3 mL, 2 mL, 1 mL, 0.7 mL, 0.5 mL, 0.4 mL, 0.3 mL, 250μL, 200 μL, 175 μL, 160 μL, 150 μL, 140 μL, 130 μL, 120 μL, 110 μL, 100μL, 70 μL, 50 μL, 30 μL, 20 μL, 10 μL, 7 μL, 5 μL, 3 μL, 1 μL, 500 nL,300 nL, 100 nL, 50 nL, 10 nL, 5 nL, 1 nL, 500 pL, 100 pL, 50 pL, 10 pL,5 pL, 1 pL, or any other volume described elsewhere herein. The pipettemay be capable of dispensing no more than, and/or equal to any fluidvolume, such as those as described herein, while having any dimension,such as those described elsewhere herein. In one example, a fluidhandling apparatus may have a height, width, and/or length that does notexceed 20 cm and a pipette head which may be capable of aspiratingand/or dispensing at least 150 μL.

The fluid handling system may be able to dispense and/or aspirate fluidwith great precision and/or accuracy. For example, coefficient ofvariation of the fluid handling system may be less than or equal to 20%,15%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1.5%, 1%, 0.7%, 0.5%,0.4%, 0.3%, 0.2%, 0.1%, 0.07%, 0.05%, 0.01%, 0.005%, or 0.001%. A fluidhandling apparatus may be capable of dispensing and/or aspirating afluid while functioning with a coefficient of variation value asdescribed herein. The fluid handling system may be able to control thevolume of fluid dispensed to within 5 mL, 3 mL, 2 mL, 1 mL, 0.7 mL, 0.5mL, 0.3 mL, 0.1 mL, 70 μL, 50 μL, 30 μL, 20 μL, 10 μL, 7 μL, 5 μL, 3 μL,1 μL, 500 nL, 300 nL, 100 nL, 50 nL, 10 nL, 5 nL, 1 nL, 500 pL, 100 pL,50 pL, 10 pL, 5 pL, or 1 pL. For example, the fluid handling apparatusmay be capable of dispensing and/or aspirating a minimum increment of nomore than any of the volumes described herein.

The fluid handling system may be capable of operating with any of thecoefficient of variations described herein and/or controlling the volumeof fluid dispensed to any value described herein while having one ormore other feature described (e.g., having any of the dimensionsdescribed herein or being capable of dispensing and/or aspirating anyvolume described herein). For example, a fluid handling apparatus may becapable of dispensing and/or aspirating 1 μL-3 mL of fluid whilefunctioning with a coefficient of variation of 4% or less.

A fluid handling apparatus may include one pipette head or a pluralityof pipette heads. In some embodiments, the plurality of pipette headsmay include a first pipette head and a second pipette head. The firstand second pipette heads may be capable of and/or configured fordispensing and/or aspirating the same amount of fluid. Alternatively,the first and second pipette heads may be capable of and/or configuredfor dispensing different amounts of fluid. For example, the firstpipette head may be configured to dispense and/or aspirate up to a firstvolume of fluid, and the second pipette head may be configured todispense and/or aspirate up to a second volume of fluid, wherein thefirst and second volumes are different or the same. In one example, thefirst volume may be about 1 mL, while the second volume may be about 250μL.

In another example, the fluid handling apparatus may include a pluralityof pipette heads, wherein a first pipette head may comprise a firstpipette nozzle configured to connect with a first removable tip, and asecond pipette head may comprise a second pipette nozzle configured toconnect with a second removable tip. The first removable tip may beconfigured to hold up to a first volume of fluid, and the secondremovable tip may be configured to hold up to a second volume of fluid.The first and second volumes may be the same or may be different. Thefirst and second volumes may have any value as described elsewhereherein. For example, the first volume may be about 1 mL, while thesecond volume may be about 250 μL.

A plurality of pipette heads may be provided for a fluid handlingapparatus. The plurality of pipette heads may be any distance apart. Insome embodiments, the fluid handling apparatus may be less than or equalto about 0.1 mm, 0.3 mm, 0.5 mm, 0.7 mm, 1 mm, 1.5 mm, 2 mm, 2.5 mm, 3mm, 3.5 mm, 4 mm, 4.5 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 10 mm, 12 mm, 15mm, 20 mm, 30 mm, or 50 mm. The distance between the pipette heads maybe from center to center of the pipette heads. The distance between thepipette heads from center to center may be the pitch of the pipetteheads.

The pipette heads may share a support structure. In some embodiments,the support structure may be a movable support structure. One, two ormore pipette heads may be movable along the support structure so thatthe lateral distance between the pipette heads may be variable. In someinstances, the pitch of the pipette heads may be variable to encompassor be limited by one or more of the dimensions previously described. Inone example, the pipette heads may be slidable along the support so thatthe distances from center to center of the pipette heads may vary. Eachof the pipette heads may be variable so that they are the same distanceapart, or may be individually variable so that they may be at variousdistances apart. A lateral distance proportion between the pipette headsmay remain the same as pipette head positions vary, or may change.Pipettes, blades, or nozzles may change their relative position (move inor out, expand or shrink) to achieve different pitches as needed and mayaccess resources in multiple planes at one time.

In some embodiments, the form factors of pipettes (e.g., positivedisplacement pipette, suction-type pipette, air displacement pipette)may be suitable for so-called “mini” pipettes. The form factors in suchcases may be reduced and optimized for space through horizontal orclamshell configurations. Systems or devices may include one or aplurality of mini pipettes. The mini pipettes may be modular andremovable from supporting structures having the mini pipettes.

In some embodiments, a mini pipette is configured to handle a sample of1 uL, 0.9 uL, 0.8 uL, 0.7 uL, 0.6 uL, 0.5 uL, 0.4 uL, 0.3 uL, 0.2 uL,0.1 uL, 10 nL, 1 nL.

In some embodiments, a mini pipette is provided that enables macro-scaleprotocol and/or processing of various chemistries at a point of servicelocation as opposed to microfluidic-restricted processing, which may notreplicate lab protocols. In some situations, the protocol and/orprocessing is selected from, without limitation: centrifugation,separation, precipitation, denaturation, extraction, coacervation,flocculation, chromatography, column based processing, elutions,dilutions, mixing, incubations, cell lysis, fixation of cells, heating,cooling, distribution of sample, separate processing or assay ordetection systems, modularity, efficiency of sample utilization,sedimentation, concentration of analyte on solid phase, immunoassay,nucleic acid amplification, nuclear magnetic resonance, microscopy,spectrometry, calorimetry, sequencing, pathological oversight andanalyses, and culture.

Pipette Configuration

A fluid handling apparatus may be a pipette. In some embodiments, afluid handling apparatus may comprise one or more pipette head. A fluidhandling apparatus may include a supporting body, and extendingtherefrom, the one or more pipette heads. As previously described, thesupporting body may support the weight of the one or more pipette heads.The supporting body may contain mechanisms for moving the pipette headsindependently or together in one dimension or multiple dimensions. Thesupporting body may permit the pipette heads to move together. Thesupporting body may also be flexible or “snake-like” and/or robotic innature, permitting the pipette heads a wide range of movement inmultiple (or infinite) directional planes. This range of movement maypermit the pipettes to serve as robotic end effectors for the devicewith one or more fluid handling or non-fluid handling functions. Thesupporting body may connect the pipette heads to one another. The sharedsupporting body may or may not be integrally formed with the pipetteheads. The supporting body may or may not also support an actuationmechanism. The supporting body may or may not be capable of supportingthe weight of actuation mechanism that may be operably connected to oneor more pipette head.

A pipette head may comprise a pipette nozzle configured to connect witha removable tip. The pipette head may also include a pipette body. Thepipette nozzle may be coaxial with the pipette body. The pipette bodymay support the pipette nozzle. The pipette nozzle may include anopening. The pipette head may also include a fluid path therein. Thefluid path may or may not be contained within the pipette body. Thefluid path may pass through the pipette body. The fluid path may have agiven length. The fluid path may terminate at the pipette nozzle. Thefluid path may be within an inner tubing. The inner tubing may be rigidor flexible.

The pipette nozzle may connect with the removable tip in any manner. Forexample, the pipette nozzle may connect with the removable tip to form afluid-tight seal. The removable tip may be friction-fit with the pipettenozzle. The tip may interface with the pipette nozzle along an outersurface of the pipette nozzle, inner surface of the pipette nozzle, orwithin a groove or intermediate portion of the pipette nozzle.Alternatively, the pipette nozzle may interface with the tip along theouter surface of the tip, inner surface of the tip, or within a grooveor intermediate portion of the tip.

In some embodiments, a plunger may be provided within a pipette head.The plunger may permit the dispensing and/or aspiration of fluid. Theplunger may be movable within the pipette head. The pipette may becapable of loading the desired plunger from a selection of plungers,that are either stored in the pipette or picked up from a storage areaoutside the pipette. The plunger may be movable along a fluid path. Theplunger may remain in the same orientation, or may have varyingorientations. In alternate embodiments, a transducer-driven diaphragmmay be provided which may affect a fluid to be dispensed and/oraspirated through the tip. Alternate dispensing and/or aspirationmechanisms may be used, which may include a positive and/or negativepressure source that may be coupled to a fluid path.

In some embodiments, the tip of the pipette head may have a length. Thedirection of tip may be along the length of the tip. In someembodiments, the fluid handling apparatus may include a motor having arotor and stator. The rotor may be configured to rotate about an axis ofrotation. The axis of rotation may have any orientation with respect tothe tip. For example, the axis of rotation may be substantially parallelto the tip. Alternatively, the axis of rotation may be substantiallynon-parallel to the tip. In some instances, the axis of rotation may besubstantially perpendicular to the tip, or any other angle with respectto the tip including but not limited to 15 degrees, 30 degrees, 45degrees, 60 degrees, or 75 degrees. In one example, the axis of rotationmay be horizontal, while the removable tip may be aligned vertically.Alternatively, the axis of rotation may be vertical while the removabletip is aligned horizontally. This configuration may provide a “bent”pipette configuration where the tip is bent relative to the motor. Themotor may be useful for metering fluid within the tip. In someembodiments, the motor may permit the movement of one or more plungerwithin a pipette head.

In some embodiments, the fluid handling apparatus may include a motorthat may be capable of permitting the movement of a plurality ofplungers that are not substantially parallel to the removable tip.Alternatively, the movement of the plurality of plungers may besubstantially parallel to the removable tip. In some instances, themovement of the plurality of plungers may be substantially perpendicularto the removable tip, or any other angle, including but not limited tothose mentioned elsewhere herein. In one example, the plunger may becapable of moving in a horizontal direction, while the removable tip isaligned vertically. Alternatively, the plunger may be capable of movingin a vertical direction while the removable tip is aligned horizontally.

A fluid path may terminate at a pipette nozzle. In some instances,another terminus of the fluid path may be provided at the plunger. Insome embodiments, the fluid path may be bent or curved. A first portionof a fluid path may have a different orientation than a second portionof the fluid path. The first and second portions may be substantiallyperpendicular to one another. The angles of the first and secondportions may be fixed relative to one another, or may be variable.

Actuation

A fluid handling apparatus may include an actuation mechanism. In someembodiments, a single actuation mechanism may be provided for the fluidhandling apparatus. Alternatively, a plurality of actuation mechanismsmay be provided. In some instances, only a single actuation mechanismmay be provided per particular use (e.g., tip removal, plunger control,switch control). Alternatively, multiple actuation mechanisms may beprovided for a particular use. In one example, an actuation mechanismmay be a motor. The motor may include a rotor and stator. The rotor maybe capable of rotating about an axis of rotation.

A single actuation mechanism, such as a motor, may be useful forindividualized dispensing and/or aspiration. A fluid handling apparatusmay include a plurality of pipette heads. A plurality of plungers may beprovided, wherein at least one plunger may be within a pipette head andconfigurable to be movable within the pipette head. In some instances,each of the pipette heads may have one or more plungers therein. Theplurality of plungers may be independently movable. In some instances,the plungers may move along a fluid path within the pipette head. Theactuation mechanism may be operably connected to the plungers. Theactuation mechanism may permit the independent movement of the pluralityof plungers. The movement of such plungers may optionally cause thedispensing and/or aspiration of fluid. A single motor or other actuationmechanism may control the independent movement of a plurality ofplungers. In some instances, a single motor or other actuation mechanismmay control the independent movement of all of the plungers within saidplurality.

A single actuation mechanism, such as a motor, may be useful forindividualized removal of a tip from pipette nozzle. A fluid handlingapparatus may include a plurality of pipette heads. A plurality of tipremoval mechanisms may be provided, wherein at least one tip removalmechanism is configured to remove an individually selected tip from thepipette nozzle. The tip removal mechanism may be configured to bemovable with respect to the pipette nozzle to effect said removal. Thetip removal mechanisms may be independently movable. Alternatively, thetip removal mechanisms need not move, but may be independentlycontrollable to permit the removal of the tips. The actuation mechanismmay be operably connected to the tip removal mechanisms. The actuationmechanism may permit the independent movement of the plurality of tipremoval mechanisms. A single motor or other actuation mechanism maycontrol the independent movement of a plurality of tip removalmechanisms. In some instances, a single motor or other actuationmechanism may control the independent movement of all of the tip removalmechanisms within said plurality.

In some embodiments, a tip removal mechanism may be within a pipettehead. An internal tip removal mechanism may be configured to be movablewithin the pipette head. For example, a tip removal mechanism may be aplunger. In other embodiments, the tip removal mechanism may be externalto the pipette head. For example, the tip removal mechanism may be acollar wrapping around at least a portion of a pipette head. The collarmay contact a portion of the pipette nozzle, pipette body and/or pipettetip. Another example of an external removal mechanism may be a strippingplate. A tip removal mechanism may or may not contact the tip whencausing the tip to be removed from the pipette.

A single actuation mechanism, such as a motor, may be useful forindividualized retraction and/or extension of a pipette nozzle. A fluidhandling apparatus may include a plurality of pipette heads. A pipettehead may include a pipette nozzle which may or may not be movable withrespect to a support body. A plurality of pipette nozzles may beindependently movable. The actuation mechanism may be operably connectedto the pipette nozzles or other portions of a pipette head that maypermit the retraction and/or extension of a pipette nozzle. Theactuation mechanism may permit the independent movement of the pluralityof pipette nozzles. A single motor or other actuation mechanism maycontrol the independent movement of a plurality of pipette nozzles. Insome instances, a single motor or other actuation mechanism may controlthe independent movement of all of the pipette nozzles within saidplurality.

In some embodiments, a tip may be connected to a pipette nozzle based onthe positions of the pipette nozzles. For example, a pipette nozzle maybe extended and brought down to contact a tip. The pipette nozzle andtip may be press-fit to one another. If selected pipette nozzles areindependently controllable to be in an extended position, the tipsconnected to the apparatus may be controllable. For example, one or morepipette nozzle may be selected to be extended. Tips may be connected tothe extended pipette nozzle. Thus, a single actuation mechanism maypermit the independent selection and connection/pick-up of tips.

Alternatively, a single motor or other actuation mechanism may controlthe independent movement of a single plunger, tip removal mechanism,and/or pipette nozzle. In some instances, a plurality of actuationmechanisms may be provided to control the movement of a plurality ofplungers, tip removal mechanisms, and/or pipette nozzles.

A fluid handling apparatus may include one or more switches. Anindividual switch may have an on position and an off position, whereinthe on position may permit an action or movement in response to movementby an actuation mechanism, and wherein the off position does not permitan action or movement in response to movement by the actuationmechanism. An on position of a switch may permit an operable connectionbetween the actuation mechanism, and another portion of the fluidhandling apparatus, such as a plunger, tip removal mechanism, and/orpipette nozzle movement mechanism. An off position of a switch may notpermit an operable connection between the actuation mechanism, andanother portion of the fluid handling apparatus, such as a plunger, tipremoval mechanism, and/or pipette nozzle movement mechanism. Forexample, an off position may permit the actuation mechanism to move, butno response is provided by the selected other portion of the fluidhandling mechanism. In one example, when a switch is in an on position,one or more plunger associated with the individual switch may move inresponse to a movement by a motor, and when the switch is in an offposition, one or more plunger associated with the individual switch isnot permitted to move in response to movement by the motor. In anotherexample, when a switch is in an on position, one or more tip removalmechanism associated with the individual switch may cause a tip to beremoved in response to movement by a motor, and when the switch is in anoff position, one or more tip removal mechanism may not cause a tip tobe removed in response to movement by the motor. Similarly, when aswitch is in an on position, one or more pipette nozzle associated withthe individual switch may extend and/or retract in response to amovement by a motor, and when the switch is in an off position, one ormore pipette nozzle associated with the individual switch is notpermitted to extend and/or retract in response to movement by the motor.

A switch may be a binary switch that may have only an on position and anoff position. One, two or more actuations may occur when a switch is inan on position and may not occur when a switch is in an off position. Inalternate embodiments, a switch may have multiple positions (e.g.,three, four, five, six, seven, eight or more positions). A switch may becompletely off, completely on, or partially on. In some embodiments, aswitch may have different degrees of depression. Different positions ofthe switch may or may not permit different combinations of actuation. Inone example, a switch in a zero position may not permit actuation of aplunger and of a tip removal mechanism, a switch in a one position maypermit actuation of a plunger while not permitting actuation of a tipremoval mechanism, a switch in a two position may not permit actuationof a plunger while permitting actuation of a tip removal mechanism, anda switch in a three position may permit actuation of a plunger andpermit actuation of a tip removal mechanism, when a motor is actuated.In some embodiments, a switch may include a control pin which may extendvarying degrees to represent different positions of the switch.

In some embodiments, the switch may be a solenoid. The solenoid may havean on position and/or an off position. In some embodiments, the solenoidmay have an extended component for an on position, and a retractedcomponent for an off position. A single solenoid may be provided foreach pipette head. For example, a single solenoid may or may not permitthe movement of an individual plunger associated with the solenoid, atip removal mechanism associated with the solenoid, or a pipette nozzleassociated with the solenoid.

Another example of a switch may include the use of one or more binarycams. FIG. 54 shows an example of a cam-switch arrangement. A cam-switcharrangement may include a plurality of binary cams 5410 a, 5410 b, 5410c, 5410 d. The binary cams may have one or more protruding segments 5420and one or more indented segments 5422. One or more control pin 5430 maybe provided. In some embodiments, each cam may have a control pinoperably connected thereto.

An individual control pin 5430 may contact an individual binary cam5410. In some embodiments, a biasing force may be provided on thecontrol pin that may cause it to remain in contact with a surface of thecam. Thus, a control pin may contact a protruding segment 5420 of thecam or an indented segment 5422 of the cam. A cam may rotate, causingthe portion of the cam contacting the control pin to change. The cam mayhave an axis of rotation. As the cam rotates, the control pin maycontact a protruding segment or an indented segment, which may cause thecontrol pin to move in response. When a control pin contacts aprotruding segment, the control pin may extend further from the axis ofrotation of the cam, than if the control pin was contacting an indentedsegment.

A plurality of cams may be provided. In one example, each of the camsmay share an axis of rotation. In some instances, the cams may have acommon shaft. The cams may be configured to rotate at the same rate. Thecams may have protruding and indented segments at different degreesabout the cam. For example, FIG. 54A shows a first cam 5410 a having oneprotruding segment, and one indented segment. A second cam 5410 b mayhave two protruding segments and two indented segments. A third cam 5410c may have four protruding segments and four indented segments. A fourthcam 5410 d may have eight protruding segments and eight indentedsegments. In some instances, any number of cams may be provided. Forinstances, n cams may be provided, where n is any positive whole number.A first cam through nth cam may be provided. Any selected cam i amongthe plurality of cams may be provided. In some instances, the ith cammay have 2^(i-1) protruding segments, and 2^(i-1) indented segments. Theprotruding and indented segments may be radially evenly spaced aroundthe cam. The configurations of the control pins that may or may notprotrude from the cams may form a binary configuration.

FIG. 54A shows an example of a binary cam at zero position, with the camrotated 0 degrees. Each of the control pins is contacting an indentedportion, which permits each of the control pins to have a retractedposition. FIG. 54B shows an example of a binary cam at one position,with the cam rotated 22.5 degrees. Each of the control pins except thefourth control pin is contacting an indented portion. The fourth controlpin is contacting a protruding segment, which causes the fourth controlpin to extend. A binary reading may be made where the retracted pins arezero, and the extended pin is 1. FIG. 54C shows an example of a binarycam at five position, with the cam rotated 112.5 degrees. The first andthird control pins are contacting an indented portion, while the secondand fourth pins are contacting a protruding portion. The second andfourth pins are extended. FIG. 54D shows an example of a binary cam atfifteen position, with the cam rotated 337.5 degrees. Each of thecontrol pins is contacting a protruding segment of the cam. Each of thecontrol pins are at an extended position, thus each having a readingof 1. The cams may be rotated any amount, which may permit anycombination of pins being extended or retracted.

An extended control pin may permit an operable connection between anactuation mechanism and another portion of the fluid handling apparatus.For example, an extended control pin for a particular cam may permit amotor to move a plunger, tip removal mechanism, and/or pipette nozzleassociated with that individual cam.

FIG. 54E shows a selection cam mounted with a motor in accordance withan embodiment of the invention. One or more cams 5410 may be providedwith one or more control pins 5430. The cams may share a shaft 5440. Amotor 5450 with an encoder may be provided. A pulley 5460 may operablyconnect the motor to the cams. In some embodiments, a motor may becapable of rotating, which may cause the cams to rotate. The shaft mayrotate, which may cause the cams to rotate together. The cams may berotated to a desired position to provide a desired arrangement ofextended control pins. The extended control pins may permit an operableconnection between another motor and another portion of the pipette. Astripped pipette body 5470 may also be provided. In some embodiments, anextended control pin may be a switch in an on position, and a retractedcontrol pin may be a switch in an off position, or vice versa.

In some embodiments, aspiration and dispensing are controlledindependently from one another. This may be accomplished with the aid ofindividual actuation mechanisms. In an example, an actuation mechanismprovides sample (e.g., fluid) aspiration while another actuationmechanism provides sample dispensing.

Venting

One or more fluid handling mechanism may include a vent. For example, apipette may include a vent. For example, a pipette nozzle and/or pipettetip may include a ventilation opening. A ventilation opening may permitan internal plunger mechanism to move within without expelling oraspirating fluid. In some embodiments, the ventilation opening maypermit a plunger to move without causing fluid within a fluid path tomove substantially along the fluid path. For example, the vent may becapable of permitting a plunger to move down within the pipette nozzleor tip without expelling the fluid. The plunger may or may not evercontact the fluid. In some instances, the plunger may move down withoutexpelling fluid until the plunger contacts the fluid. In anotherexample, a ventilation opening may permit a plunger to move upwards awayfrom a fluid and draw in air, while permitting the fluid to remain inits position within the pipette nozzle or tip.

A vent may permit increased accuracy and/or precision of a pipette. Thevent may be included in air displacement pipettes. The vent may increasethe accuracy and/or precision of an air displacement pipette bypermitting the venting of air that may cause inherent inaccuracies withthe fluid, depending on environmental conditions. Alternatively, thevent may be included for positive displacement pipettes. Venting mayreduce inaccuracies associated with variable conditions. The vent maypermit pipette tips filled with fluid to be ejected without loss offluid from the tips. Venting fluid-filled tips without loss of fluid mayenable incubation of tips when disengaged from the pipette, therebyfreeing up the pipette to execute other tasks. In an embodiment, thepipette tips may be vented, and later picked up for further processingof the fluid inside.

In some embodiments, a fluid handling apparatus may include one or moreventilation port. In some instances, one or more pipette head may have aventilation port. In one example, each pipette head of the fluidhandling apparatus may have a ventilation port. Each pipette head of aparticular type (e.g., air displacement pipette head) may have aventilation port.

A ventilation port may be capable of having an open position and aclosed position. In some instances, a switch may be used to determinewhether the ventilation port is in an open position or a closedposition. In one example, the switch may be a solenoid, valve, or anyother switching mechanism described elsewhere herein. The ventilationport switch may have one or more characteristic provided for any otherswitching mechanism described elsewhere herein, or vice versa. Theventilation port switch may be a binary switch, or may have multiplepositions. A ventilation port may either be open or closed, or may havevarying degrees of openness. Whether the ventilation port is open orclosed, or the degrees of openness of the ventilation port may becontrolled by a controller. In one example, a ventilation solenoid maydetermine whether the ventilation port is in an open position or closedposition. In another example, a valve may determine whether theventilation port is in the open position or closed position. A valve,solenoid, or any other switch may be duty cycled. The duty cycling mayhave any period, including but not limited to periods of 5 s or less, 3s or less, 2 s or less, 1 s or less, 500 ms or less, 300 ms or less, 200ms or less, 100 ms or less, 75 ms or less, 60 ms or less, 50 ms or less,40 ms or less, 30 ms or less, 20 ms or less, 10 ms or less, 5 ms orless, or 1 ms or less. The duty cycle may be controlled in accordancewith one or more instructions from a controller.

In some embodiments, a ventilation solenoid, valve, or other switch maydetermine the degree to which a vent may be opened. For example, theswitch may only determine if the ventilation port is open or closed.Alternatively, the switch may determine whether the ventilation port isopen to an intermediary degree, such as about 10%, 20%, 30%, 40%, 50%,60%, 70%, 80%, or 90% open. The ventilation port may be open to a fixeddegree, or may open any degree along a continuous spectrum. The degreeof opening may be controlled in response to one or more signal from acontroller. The controller may be used to determine a desired degree ofpressure to be provided in a fluid path.

A ventilation port may be coupled to a pressure source. The pressuresource may be a positive pressure source or a negative pressure source.The positive pressure source may be useful for expulsion of a fluid fromwithin the pipette head. The negative pressure source may be useful forthe aspiration of fluid into the pipette head. In some instances, theventilation port may be coupled to atmospheric conditions. Forinstances, the ventilation port may selectively connect an interior ofthe pipette head with ambient air.

The positive or negative pressure may be delivered by any techniqueknown in the art. In one example, the ventilation port may be coupled toa reversible pump capable of delivering positive or negative pressure.The pump may be capable of delivering the positive or negative pressurefor an extended period of time. For example, the pump may deliver thepositive pressure until all fluid is expelled. The pump may deliver thepositive pressure as long as desired in order to permit a desired amountof fluid to be expelled through the pipette head. In another example,the pump may deliver a negative pressure as long as desired in order topermit the desired amount of fluid to be aspirated through the pipettehead. The reversible pump may permit switching between providingpositive and negative pressure under selected conditions.

The positive or negative pressure may be provided by a fluid. Forexample, the positive or negative pressure may be provided by air oranother gas. In other embodiments, the positive or negative pressure maybe provided by liquid, or any other fluid.

In some instances, a pipette head has a single ventilation port.Alternatively, a pipette head may have multiple ventilation ports.Multiple ventilation ports may be connected to positive pressuresources, negative pressure sources, ambient conditions, or anycombinations thereof.

Retraction

A fluid handling apparatus may include one or more pipette head, whereinan individual pipette head has a fluid path of a given length. The fluidpath may be entirely within the pipette head, or one or more portion ofthe pipette head may be outside the pipette head. The fluid path lengthmay terminate at a pipette nozzle. The fluid path length may terminateat an orifice of the fluid handling apparatus. In some instances, thefluid path length may terminate at an end of a tip connected to thefluid handling apparatus. In some instances, a fluid path length mayterminate at the end of a plunger (e.g., the end of the plunger closerto the tip). Alternatively, the fluid path length may terminate at anend of a pipette head or base or support. The fluid path may have two ormore termination ends, which may be any combination of the terminationlocations mentioned above. In some instances, the fluid path length maybe determined by two termination ends.

The length of the fluid path may be adjustable. In some instances, thelength of the fluid path may be adjustable without effecting movement offluid from a tip, when the tip and pipette nozzle are engaged. The fluidpath length may be adjusted while the fluid within a tip remains atsubstantially the same position. The fluid path length may be increasedand/or decreased.

The fluid path length may be adjusted by altering the position of one,two, or more of the termination points of the fluid path. In oneexample, a fluid path may have two termination points, a distaltermination point that is closer to the tip or the point at which fluidis expelled and/or aspirated, and a proximal termination point that isfurther from the tip or the point at which fluid is expelled and/oraspirated. A distal termination point may be moved, thereby adjustingthe fluid path length. Alternatively, a proximate termination point maybe moved, thereby adjusting the fluid path length. In some instances,the distal and proximal termination points may be moved relative to oneanother, thereby adjusting the fluid path length.

In one example, a distal termination point may be a pipette nozzle, anda proximal termination point may be a plunger end closer to the pipettenozzle. The pipette nozzle may be connected to a tip which may contain afluid therein. The pipette nozzle may be retracted or extended relativeto the plunger and/or the rest of the pipette head. The fluid pathlength of the pipette head may be adjusted. In some instances, extendingand/or retracting the pipette nozzle need not cause substantial movementof the fluid within the tip. In another example, the plunger may beactuated toward or away from the tip. This may also cause fluid pathlength of the pipette head to be adjusted. The plunger may be actuatedwithout causing substantial movement of the fluid within the tip.

As previously described, a fluid handling apparatus may include at leastone pipette head connected to a base, wherein an individual pipette headcomprises a pipette nozzle configured to connect with a removable tip. Aplunger may be provided within the pipette head, and may be configuredto be movable within the pipette head. The pipette nozzle may be movablerelative to the base, such that the pipette nozzle is capable of havinga retracted position and an extended position, wherein the pipettenozzle is further away from the base than in the retracted position. Thepipette nozzle may be movable relative to the plunger, to the motor, tothe rest of the pipette head, to the switch, or to any other portion ofthe fluid handling apparatus. Adjusting the pipette nozzle between theretracted and extended position may change a fluid path lengthterminating at the pipette nozzle. In some instances, the fluid pathlength may be formed using only rigid components.

Any difference in position may be provided between the retractedposition and the extended position. For example, no more than and/orequal to about a 1 mm, 3 mm, 5 mm, 7 mm, 1 cm, 1.5 cm, 2 cm, 2.5 cm, 3cm, 4 cm, 5 cm, or 10 cm difference may exist between the retractedposition and the extended position. The difference in position may be ina vertical direction, horizontal direction, or any combination thereof.The difference in position may be in a direction parallel to the lengthof the tip, perpendicular to the length of the tip, or any combinationthereof.

In some embodiments, this may be enabled by venting, such as ventilationmechanisms described elsewhere herein, or other mechanisms. Theventilation port may be located along the fluid path.

The fluid path may be formed from one or more components. In someembodiments, the fluid path may be formed entirely of rigid components.In other embodiments, the fluid path may be formed from flexiblecomponents. Alternatively, the fluid path may be formed from acombination of rigid and flexible components. The fluid path may beformed from rigid components without the use of flexible components. Thefluid path may be formed from flexible components without the use ofrigid components.

Examples of rigid components may include hard tubes, pipes, conduits, orchannels. The fluid path may be formed from a single rigid component ormultiple rigid components. Multiple rigid components may or may not bemovable relative to one another. The rigid components may slide relativeto one another. In one example, a plurality of rigid components may beprovided in a telescoping configuration, where one or more rigidcomponent may slide within another rigid component. The length of thefluid path may be altered by moving the one or more rigid componentsrelative to one another.

Examples of flexible components may include bendable tubes, pipes,conduits or channels. For example, bendable plastic tubing may be used.The fluid path may be formed from a single flexible component ormultiple flexible components. Multiple flexible components may bemovable relative to one another. For instance, they may slide relativeto one another, and/or may have a telescoping arrangement.

A fluid handling apparatus may have a plunger within one or more pipettehead. The plunger may be configured to be movable within the pipettehead. The plunger may be movable along a fluid path. The plunger may bemovable in a vertical direction and/or a horizontal direction. Theplunger may be movable in a direction parallel to the length of a tipand/or perpendicular to the length of the tip. The plunger may form afluid-tight connection with one or more walls of the fluid path. Thus,as the plunger may move along a fluid path, the pressure within thefluid path may be altered and/or maintained.

The plunger may be formed from rigid components, flexible components, orany combination thereof. The plunger may be formed from a singleintegral piece. Alternatively, the plunger may be formed from multiplesections. For example, the plunger may comprise a first section and asecond section. At least a portion of the first section may beconfigured to slide relative to the second section, thereby permittingthe plunger to extend and/or collapse. In one example, the first sectionmay be configured to slide within the second section. A telescopingarrangement may be provided. The length of the plunger may be fixed ormay be variable. The plunger may have any number of sections (e.g., one,two, three, four, five, six, seven, eight, or more sections), which mayor may not be movable relative to one another. The plunger may form adouble needle and/or multi-needle configuration.

In some embodiments, a heat spreader may surround the plunger. The heatspreader may assist with keeping the plunger at a desired temperature,or within a desired temperature range. This may be beneficial whenprecise control of volumes dispensed and/or aspirated is desired. Theheat spreader may assist with reducing and/or controlling thermalexpansion of one or more components of the fluid handling apparatus,such as the plunger. In other embodiments, the pipette nozzles and/ortips can be used to transfer heat to and/or from the pipette for heatingand/or cooling operations. The pipette can also be used to deliver/applycool air for controlling temperature of cartridge, vessels, tips, etc. Apump may be utilized for this function.

An aspect of the invention may be directed to a method of fluidhandling, which may include providing a fluid handling apparatus havingone or more of the features described herein. For example, the methodmay include providing at least one pipette head operably connected to abase, wherein an individual pipette head comprises a pipette nozzleconfigured to connect with a removable tip. The method may also includeretracting and/or extending the pipette nozzle relative to the base. Themethod may include retracting and/or extending the pipette nozzle anydistance, which may be dictated by a controller.

The method may optionally include dispensing and/or aspirating a fluidwith a tip. The aspirating and/or dispensing may occur while the pipettenozzle is retracting and/or extending. The aspirating and/or dispensingmay occur while the pipette nozzle is retracting and/or extending in avertical direction, horizontal direction, direction parallel to a tiplength, direction perpendicular to a tip length, away/towards a base, orany combination thereof.

The speed of dispensing and/or aspiration may depend on the speed ofretracting and/or extending by the pipette nozzle, or vice versa.Dispensing and/or aspirating during retracting and/or extending thepipette nozzle may be beneficial in systems with small volumes of fluidand small vessels. For example, a small vessel may be provided with afluid at or near the top level of the vessel. When a tip encounters thetop of the fluid surface at the vessel, if no aspirating occurs,overflow may occur. If aspiration occurs while the tip is encounteringthe fluid and lowered into the vessel, the aspirating may prevent theoverflow from occurring. In some embodiments, dispensing and/oraspirating may occur at a rate sufficient to prevent overflow, or tohave any other desirable effects.

In some embodiments, a pipette nozzle may be extended and/or retractedprior to, concurrently with, and/or subsequent to translating a pipettehead. The pipette nozzle may be extended and/or retracted in a firstdirection, and the pipette head translation may occur in a seconddirection. The first and second directions may or may not besubstantially parallel to one another. In some instances, the first andsecond directions may be substantially non-parallel to one another. Thefirst and second directions may be substantially perpendicular to oneanother. In one example, the first direction is a substantially verticaldirection while the second direction is a substantially horizontaldirection. In another example, the first direction is substantiallyparallel with the length of the tip, and the second direction issubstantially perpendicular to the length of the tip.

The pipette nozzle may be extended and/or retracted relative to the baseprior to, currently with, and/or subsequent to dispensing and/oraspirating the fluid with the tip. The fluid may be dispensed and/oraspirated prior to, currently with, and/or subsequently to translatingthe pipette head.

In one example, a pipette nozzle may be retracted prior to and/orcurrently with translating the pipette head. The pipette nozzle may thenbe extended prior to and/or concurrently with dispensing and/oraspirating a fluid with the tip. The pipette tip may be retracted asufficient amount to clear any objects that may be encountered whiletranslating the pipette head. The pipette tip may be extendedsufficiently to make contact with a fluid to be aspirated, and/or todispense the fluid to a designated location.

The pipette nozzle may or may not extend and/or retract while thetranslation of the pipette head occurs. In some instances, individualpipette nozzles of a plurality of pipette heads that are translatedtogether may or may not extend and/or retract together. In someinstances, the individual pipette nozzles may be independently retractedand/or extended. The pipette nozzle may extend and/or retract based on aknown path to be traveled, which may or may not include known obstaclesto be cleared. The pipette nozzle may extend and/or retract based on oneor more measurement provided by a sensor (e.g., if a sensor encountersan obstruction during the translation of the pipette heads).

In some situations, a pipette may include one or more sensors forproviding various data to a control system operating the pipette. In anexample, the one or more sensors provide position measurements thatenable the pipette to extend and retract. In another example, the one ormore sensors provide temperature, pressure, humidity, conductivity data.In another example, the one or more sensors include cameras for takingimage, video and/or sound recording from within the pipette.

A multi-head pipette may have a plurality of pipette heads. One or moreof the pipette heads and/or each of the pipette heads may include apipette nozzle. One or more of the pipette heads and/or each of thepipette heads may have a pipette tip connected thereto. One or more ofthe pipette heads and/or each of the pipette heads may be capable ofaccepting or connecting to a pipette tip. In one example, each pipettehead may connect to one pipette tip. In other examples, each pipettehead may be capable of connecting to one or multiple pipette tips. Thepipette tip may be press-fit onto the pipette head and/or may beconnected using any other mechanism known in the part including, but notlimited to, magnetic, snap-fit, hook and loop fasteners, elastics, ties,sliding mechanisms, locking mechanism, clamps, actuated mechanicalcomponents, and/or adhesives.

One or more of the pipette heads may be provided in a row. For example,one or more, two or more, three or more, four or more, five or more, sixor more, seven or more, eight or more, nine or more, ten or more, ortwelve or more pipette heads may be provided in a row. One or morepipette heads may be provided in a column. For example, one or more, twoor more, three or more, four or more, five or more, six or more, sevenor more, eight or more, nine or more, ten or more, or twelve or morepipette heads may be provided in a column. Arrays of pipettes may beprovided, wherein the array has one or more, two or more, three or more,four or more, five or more, six or more, seven or more, eight or more,nine or more, ten or more, or twelve or more pipette heads in the rowand one or more, two or more, three or more, four or more, five or more,six or more, seven or more, eight or more, nine or more, ten or more, ortwelve or more pipette heads in the column. In some embodiments, thepipette heads may be arranged in staggered rows, straight, curved orbent rows, concentric shapes, or any other configuration. The pipetteheads may be configured and/or dimensioned to match one or morearrangement on a microcard as described elsewhere herein.

The multi-headed pipette may have air displacement pipettes having theconfigurations of the pipette heads described elsewhere herein.Alternatively, the multi-headed pipette may have positive displacementpipettes, having the configurations of the pipette heads as describedelsewhere herein. Alternatively, the multi-headed pipette may includeboth air displacement and positive displacement pipettes. One or moreair displacement pipettes may be provided in one region and one or morepositive displacement pipette may be provided in another region.Alternatively, the air displacement pipettes and positive displacementpipette may be interspersed. The air displacement pipettes may beprovided in one format while a positive displacement pipette may beprovided in another format. For example, a row of air displacementpipettes may be provided while a single positive displacement pipettemay be provided. In one embodiment, an eight-head row of airdisplacement pipettes may be provided along with a single positivedisplacement pipette.

One or more air displacement pipette and one or more positivedisplacement pipette may be provided on the same pipette support.Alternatively, they may be provided on different pipette supports. Theair displacement pipette and positive displacement pipette may be atfixed positions relative to one another. Alternatively, they may bemovable relative to one another.

One, two, three, four, five, six or more pipettes and/or other fluidhandling mechanisms may be provided within a device. The fluid handlingmechanisms may have a fixed position within the device. Alternatively,the fluid handling mechanisms may be movable within the device.

One, two, three, four, five, six or more pipettes and/or other fluidhandling mechanisms may be provided within a module. The fluid handlingmechanisms may have a fixed position within the module. Alternatively,the fluid handling mechanism may be movable within the module. In someembodiments, the fluid handling mechanism may be movable betweenmodules. Optionally, a fluid handling mechanism may be provided externalto the modules but within the device.

The fluid handling mechanisms may transfer sample or other fluid fromone portion of the device and/or module to another. The fluid handlingmechanism may transfer fluids between modules. The fluid handlingmechanism may enable fluid to be shuttled from one portion of the deviceto another in order to affect one or more sample processing step. Forexample, a fluid may undergo a sample preparation step in a firstportion of the device, and may be transferred to a second portion of thedevice by the fluid handling system, where an additional samplepreparation step, an assay step, or a detection step may occur. Inanother example, a fluid may undergo an assay in a first portion of thedevice and may be transferred to a second portion of the device by thefluid handling system, where an additional assay step, detection step,or sample preparation step may occur. In some cases, the fluid handlingmechanism is configured to transfer a fluid, solid or semi-solid (e.g.,gel). Thus, the term “fluid handling” need not be limited to fluids, butmay capture substances of varying viscosities or consistencies.

The fluid handling may permit the transfer of fluids while the fluidsare contained within one or more pipette tips or vessels. Pipette tipsand/or vessels containing the fluid may be moved from one portion of thedevice to another. For example, a pipette tip may pick up a fluid in oneportion of the device, and be moved to a second portion of the device,where the fluid may be dispensed. Alternatively, portions of the devicemay be moved relative to the fluid handling mechanism. For example, aportion of the device may be moved to the pipette, where the pipette maypick up a fluid. Then another portion of the device may be moved to thepipette, where the pipette may dispense the fluid. Similarly, a fluidhandling mechanism may be movable to pick up and/or remove pipette tipsand/or vessels in different locations.

Fluid Handling Tips

In one example, a pipette nozzle may be configured to accept one or moretype of pipette tip. The pipette nozzle may be shaped to becomplementary to one or more type of pipette tip. In some embodiments,the pipette tips may have an end with the same diameter, even if otherpipette tip shapes or dimensions may be vary. In another example, thepipette nozzle may have one or more shaped features which mayselectively contact pipette tips depending on the pipette tip. Forexample, the pipette nozzle may have a first portion that contacts afirst type of pipette tip, and a second portion that contacts a secondtype of pipette tip. The pipette nozzles may have the same configurationin such situations. Alternatively, the pipette nozzle may be speciallyshaped to fit one type of pipette tip. Different pipette nozzles may beused for different pipette tips.

The pipette tip may be formed of a material that may enable one or moresignal to be detected from the pipette tip. For example, a pipette tipmay be transparent and may permit optical detection of fluid within thepipette tip. A pipette tip may be optically read, or detected in anyother manner while the pipette tip is attached to a pipette nozzle.Alternatively, the pipette tip may be optically read, or detected in anyother manner, when the pipette tip has been removed from the pipettenozzle. The pipette tip may or may not have a fluid contained thereinwhen read by a detector. A pipette tip may have one or moreconfiguration, dimension, characteristic, or feature as described ingreater detail elsewhere herein.

In some embodiments, a pipette tip may receive or emit a light from alight source. The tip may function as a lens to focus the light emittedby the pipette. In some embodiments, a light source may be operablyconnected to a fluid handling apparatus. The light source may beexternal to the fluid handling apparatus, or may be within the fluidhandling apparatus. In some embodiments, one or more light source may beprovided within a pipette head of the fluid handling apparatus. In someembodiments, a plurality of pipette heads or each pipette head may havea light source. A plurality of light sources may or may not beindependently controllable. One or more characteristic of the lightsource may or may not be controlled, including but not limited towhether the light source is on or off, brightness of light source,wavelength of light, intensity of light, angle of illumination, positionof light source. The light source may provide light into the tip.

A light source may be any device capable of emitting energy along theelectromagnetic spectrum. A light source may emit light along a visiblespectrum. In one example, a light source may be a light-emitting diode(LED) (e.g., gallium arsenide (GaAs) LED, aluminum gallium arsenide(AlGaAs) LED, gallium arsenide phosphide (GaAsP) LED, aluminum galliumindium phosphide (AlGaInP) LED, gallium(III) phosphide (GaP) LED, indiumgallium nitride (InGaN)/gallium(III) nitride (GaN) LED, or aluminumgallium phosphide (AlGaP) LED). In another example, a light source canbe a laser, for example a vertical cavity surface emitting laser (VCSEL)or other suitable light emitter such as anIndium-Gallium-Aluminum-Phosphide (InGaAIP) laser, a Gallium-ArsenicPhosphide/Gallium Phosphide (GaAsP/GaP) laser, or aGallium-Aluminum-Arsenide/Gallium-Aluminum-Arsenide (GaAIAs/GaAs) laser.Other examples of light sources may include but are not limited toelectron stimulated light sources (e.g., Cathodoluminescence, ElectronStimulated Luminescence (ESL light bulbs), Cathode ray tube (CRTmonitor), Nixie tube), incandescent light sources (e.g., Carbon buttonlamp, Conventional incandescent light bulbs, Halogen lamps, Globar,Nernst lamp), electroluminescent (EL) light sources (e.g.,Light-emitting diodes—Organic light-emitting diodes, Polymerlight-emitting diodes, Solid-state lighting, LED lamp,Electroluminescent sheets Electroluminescent wires), gas discharge lightsources (e.g., Fluorescent lamps, Inductive lighting, Hollow cathodelamp, Neon and argon lamps, Plasma lamps, Xenon flash lamps), orhigh-intensity discharge light sources (e.g., Carbon arc lamps, Ceramicdischarge metal halide lamps, Hydrargyrum medium-arc iodide lamps,Mercury-vapor lamps, Metal halide lamps, Sodium vapor lamps, Xenon arclamps). Alternatively, a light source may be a bioluminescent,chemiluminescent, phosphorescent, or fluorescent light source.

The light source may be capable of emitting electromagnetic waves in anyspectrum. For example, the light source may have a wavelength fallingbetween 10 nm and 100 μm. The wavelength of light may fall between 100nm to 5000 nm, 300 nm to 1000 nm, or 400 nm to 800 nm. The wavelength oflight may be less than, and/or equal to 10 nm, 100 nm, 200 nm, 300 nm,400 nm, 500 nm, 600 nm, 700 nm, 800 nm, 900 nm, 1000 nm, 1100 nm, 1200nm, 1300 nm, 1500 nm, 1750 nm, 2000 nm, 2500 nm, 3000 nm, 4000 nm, or5000 nm.

One or more of a plurality of light sources may be provided. In someembodiments, each of the plurality of light sources may be the same.Alternatively, one or more of the light sources may vary. The lightcharacteristics of the light emitted by the light sources may be thesame or may vary. The light sources may be independently controllable.

The tip may form a wave guide capable of providing light through the tipto a fluid contained therein, or capable of transmitting an opticalsignal from the fluid through the tip. The tip may be capable oftransmitting light from a light source to a fluid contained therein. Thelight source may be infrared light. The infrared light may be used toheat samples or reactions in the tip or elsewhere. The tip may becapable of transmitting light. The tip may be formed of an opticallytransmissive material. In some embodiments, the tip may transmit allwaves of the electromagnetic spectrum. Alternatively, the tip maytransmit selected waves of the electromagnetic spectrum. For example,the tip may transmit selected wavelengths of light. The tip may or maynot transmit light along the entire length of the tip. A portion or theentire tip may be formed of the optically transmissive material. The tipmay be transparent, translucent, and/or opaque.

In some embodiments, the tip may comprise a fiber that is capable ofconducting light. The fiber may be formed of an optically transparentmaterial. The fiber may extend along a portion or the entire length ofthe removable tip. The fiber optic may be embedded in the removable tip.The fiber optic may be embedded within an opaque tip, transparent tip,and/or translucent tip.

A pipette nozzle may be formed of a transparent and/or reflectivesurface. The pipette nozzle may be configured to permit the transmissionof light through the pipette nozzle. For example, light from a lightsource may pass through the pipette nozzle to the tip. In someembodiments, the pipette nozzle may have a reflective surface. Lightfrom a tip may be reflected by the pipette nozzle back into the tip,thereby creating a high degree of illumination within the tip oradjacent to the tip.

FIG. 55 shows an example of a fluid handling apparatus using one or morelight source. FIG. 55A shows a plurality of pipette heads. Each pipettehead may include a nozzle 5510. An ejection sleeve 5512 may be providedfor each pipette head.

FIG. 55B shows a side cut away view of a fluid handling apparatus with aplunger 5520 at a bottom position. The apparatus may include a pipettehousing 5530. A solenoid 5540 may be provided, which may affect theactuation of an ejection sleeve 5512 or a plunger 5520.

FIG. 55C shows a close up of a light source that may be provided withina fluid handling apparatus. For example, an LED 5550 or other lightsource may be provided within a pipette housing. Any description hereinof an LED may also apply to any other light source, and vice versa. TheLED may be located at an end of a plunger 5520. The LED may be locatedat a top end of the plunger or a bottom end of the plunger. The LED maybe coaxial with the plunger. The LED may be integral to the plunger ormay be a separate piece from the plunger. The LED may or may notdirectly contact the plunger. In some embodiments, the LED may move withthe plunger. Alternatively, the LED may remain stationary while theplunger may be movable.

A plunger holder 5560 may be provided which may assist with aligningand/or controlling the plunger position. A plunger holder may have oneor more feature 5565 which may put a plunger in an extended or retractedposition. When a plunger is in an extended position, it may be locatedcloser to a pipette nozzle, and/or tip, than when a plunger is in aretracted position.

FIG. 55D shows a close up of a plunger 5520 and pipette nozzle 5510. Insome instances, an o-ring 5570 may be provided on a pipette head. Theplunger may be formed of an optically transmissive material. In someembodiments, the plunger may be formed of a transparent material. Theplunger may be a light pipe plunger, which may function as a wave guide.The plunger may transmit light from the light source to the tip and/orfluid contained within the tip. The plunger may or may not transmitlight from a fluid within the tip to another location.

FIG. 55E shows a perspective view of a fluid handling apparatus.

A fluid handling apparatus may be operably connected to an image capturedevice. The image capture device may be capable of capturing an image ofa fluid within the tip. Alternatively, the image capture device may becapable of capturing an image through the tip. The image capture devicemay be external to the fluid handling apparatus, or may be within thefluid handling apparatus. In some embodiments, one or more image capturedevices may be provided within a pipette head of the fluid handlingapparatus. In some embodiments, a plurality of pipette heads or eachpipette head may have an image capture device. In some embodiments, theimage capture device may be integrally formed with the apparatus. Theapparatus itself may able to function as an image capture device. Insome embodiments, the tip and/or plunger may be capable of functioningas a lens of the image capture device. The tip and/or plunger may beformed of an optically transmissive material which may be shaped toprovide desirable optical effects.

A plurality of image capture devices may or may not be independentlycontrollable. The image capture devices may be the same, or may vary.

Any description of an image capture device may apply to anyelectromagnetic spectrum detecting device. The image capture device maybe capable of capturing electromagnetic emission and generating an imagealong one or more of: a visible spectrum, an infra-red spectrum, anultra-violet spectrum, or a gamma spectrum. In some embodiments, theimage capture device is a camera. Any descriptions of cameras, or otherdetection devices described elsewhere herein may apply. In one example,the image capture device may be a digital camera. Image capture devicesmay also include charge coupled devices (CCDs) or photomultipliers andphototubes, or photodetector or other detection device such as ascanning microscope, whether back-lit or forward-lit. In some instances,cameras may use CCDs, CMOS, may be lensless (computational) cameras(e.g., Frankencamera), open-source cameras, or may use any other visualdetection technology known or later developed in the art. Cameras mayinclude one or more feature that may focus the camera during use, or maycapture images that can be later focused. In some embodiments, imagingdevices may employ 2-d imaging, 3-d imaging, and/or 4-d imaging(incorporating changes over time). Imaging devices may capture staticimages. The static images may be captured at one or more point in time.The imaging devices may also capture video and/or dynamic images. Thevideo images may be captured continuously over one or more periods oftime. Any other description of imaging devices and/or detection unitsmay also be applied.

In one example, an image capture device may be located at an end of theplunger. In some examples, the image capture device may be located on abottom end or a top end of the plunger. The image capture device may becoaxial with the plunger. The image capture device may be integral tothe plunger or may be a separate piece from the plunger. The imagecapture device may or may not directly contact the plunger. In someembodiments, the image capture device may move with the plunger.Alternatively, the image capture device may remain stationary while theplunger may be movable. The image capture device may be located where alight source is located as provided in FIG. 55B and FIG. 55C, oradjacent to or in the proximity of the light source.

The plunger and/or tip may include an optically transmissive material.The plunger and/or tip may be made from a transparent material. Theplunger and/or tip may be shaped to have desirable optical properties.The plunger and/or tip may be a lens of the image capture device.Movement of the plunger and/or tip may or may not affect the focus of animage captured by the image capture device. The image capture device maybe directed in a longitudinal direction along the length of a tip.Alternatively, the image capture device may be directed in a lateraldirection perpendicular to the length of the tip, or at any other angle.

In some embodiments, the image capture device may be capable ofcapturing an image of a fluid within a tip. Alternatively, the imagecapture device may be capable of capturing an image of any sample withinthe device. In some embodiments, the image capture device may capture animage of a sample that is located at the end of a tip. For example, asample may be located at the end of a tip opposite the pipette nozzle.The image capture device may capture an image through the tip of thesample. The sample may be a fluid sample, tissue sample, or any othersample described elsewhere herein. In some embodiments, the imagecapture device may operate in conjunction with a light source. The lightsource may illuminate the sample, which may permit the image capturedevice to capture an image of the sample.

A processor may be operably connected to a tip of the fluid handlingapparatus. The processor may be located within the fluid handlingapparatus, within a pipette head associated with the tip, or on the tipitself. The fluid handling apparatus may vary and/or maintain theposition of a removable tip based on instructions from the processor.The processor may be connected to a sensor on or near the fluid handlingapparatus that measures environmental conditions (such as temperature,humidity, or vapor pressure) and may adjust the motion of the fluidhandling device to compensate or optimize for such conditions.

In one example, a plurality of tips may be provided, wherein anindividual tip of said plurality may have a processor on and/or beoperably connected to the tip. In some embodiments, each tip may have aprocessor thereon or operably connected. The tip processors may becapable of communicating with a controller and/or with one another. Forinstances, a first processor of a first removable tip may be incommunication with a second processor of a second removable tip.

In some embodiments, based on said communications, the location of thetip may be controllable. The location of the tips may be controllablewhile they are engaged with a pipette head. Alternatively, the locationof the tips may be controllable when they are separated from a pipettehead. The tips may be capable of varying and/or maintaining theirposition while they are engaged with a pipette head and/or while theyare separated from a pipette head.

A tip may include one, two, or more openings. A tip is any useful shapethat can interface with the pipette or one or more pipette nozzles. Atip can take many forms, such as cylindrical, elliptical, square,“T”-shaped, or round shapes. A single tip may have multiplesub-compartments or wells. Such sub-compartments may be used to containvarious useful chemicals, such as reagents. Useful chemicals such asreagents may be deposited in or on the tip or any of its subcompartmentsin liquid, solid, film or other form. Tips may contain vesicles ofchemicals, such as reagents, that may be released on command (e.g., whenpierced). Tips can also be used for chemical and physical processingsteps, such as filtration of reagents and/or samples. One or more of theopenings may include a switch, such as a valve. In one example, a tipmay have two openings, each of which may include an embedded passivevalve. A switch, such as an embedded passive valve may be configured topermit fluid to flow in one direction through a first opening, andthrough a tip body, and through a second opening. A valve may control adirection of fluid flow. The fluid may flow entirely through the tip, ormay flow through a portion of the tip. For example, a tip may have aswitch at one opening, which may permit fluid to flow in a certaindirection (e.g., fluid to flow into the tip to permit aspiration whilenot allowing fluid to fall out of the tip, or fluid to flow out of thetip to permit dispensing while not allowing fluid to be aspirated intothe tip. The valves may be controlled to determine the direction offluid flow, magnitude of fluid flow, or whether any fluid is permittedto flow.

The fluid handling system may be able to simultaneously dispense and/oraspirate one or a plurality of fluids. In some instances, the fluidhandling system may be dispensing, aspirating, and/or transporting aplurality of types of fluids simultaneously. The fluid handling mayprovide a modularized technique of tracking and handling differentfluids for one or more concurrent steps or tests.

Multi-Use Transport

A fluid handling apparatus may be useful to dispense, aspirate, and/ortransfer one or more fluids. The fluid handling apparatus may also beuseful for one or more additional function, including non-fluid handlingfunctions. The connection of a component or tip may permit the fluidhandling device to function as a robot capable of performing one or morenon-fluid handling functions. Alternatively, the pipette itself may beemployed to perform one or more such non-fluid handling functions bymeans of one or more actuation mechanisms. Such non-fluid handlingfunctions may include the ability to transfer power to move components,tools or other objects, such as a cuvette body, or cartridges or testsamples, or any component thereof. When combined with a flexiblesupporting body (described herein) or other configuration allowing awide range of movement, the apparatus may be able to perform suchfunctions in multiple dimensions within the device, or even outside it.

For instance, the fluid handling apparatus may be useful to transfer acomponent from one location within the device, to another. Componentsthat may be transferred may be sample processing components. A sampleprocessing component may be a sample preparation unit or componentthereof, an assay unit component thereof, and/or a detection unit orcomponent thereof. Examples of components may include but are notlimited to tips, vessels, support structures, micro cards, sensors,temperature control devices, image capture units, optics, cytometers,centrifuges, or any other components described elsewhere herein.

The fluid handling apparatus may pick up a sample processing component.The fluid handling apparatus may move the sample processing component toa different location of the device. The fluid handling apparatus maydrop off the sample processing component at its new location within thedevice.

The fluid handling apparatus may be capable of transferring sampleprocessing components within a module. The fluid handling apparatus mayor may not be confined to the module. Alternatively, the fluid handlingapparatus may be capable of transferring sample processing componentsbetween modules, and need not be confined to a single module. In someinstances, the fluid handling apparatus may be capable of transferringsample processing components within a rack and/or may be confined to arack. Alternatively, the fluid handling apparatus may be capable oftransferring sample processing components between racks, and need notconfined to a single rack.

A fluid handling apparatus may pick up and move a sample processingcomponent using various mechanisms. For example, the sample processingcomponent may be picked up using a press-fit between one or more of thepipette heads and a feature of the sample processing component. Forexample, a pipette nozzle may interface with a tip through a press-fitarrangement. The same press-fit arrangement may be used to permit apipette nozzle and a feature of the sample processing component toengage. Alternatively, the press-fit interface may occur between anyother portion of the fluid handling apparatus and the sample processingcomponent. In some instances, the press-fit feature of the sampleprocessing component may be protruding to encounter the fluid handlingapparatus. The press-fit feature of the sample processing component mayhave a shape complementary to the press-fit portion of the fluidhandling apparatus.

Another example of an interface mechanism may be a pressure-drivenmechanism, such as a suction mechanism. The sample processing componentmay be picked up using a suction provided by one, two or more of thepipette heads. The suction may be provided by one or more pipette headmay be provided by the internal actuation of a plunger, or a negativepressure source coupled to the fluid path. The pipette heads providingsuction may contact any portion of the sample processing component, ormay contact a specific feature of the sample processing component. Thefeature of the sample processing component may or may not be protrudingto encounter the fluid handling apparatus.

An additional example of an interface mechanism may be a magneticmechanism. A fluid handling apparatus may include a magnet that may beturned on to interface with a magnet of the sample processing component.The magnet may be turned off when it is desired to drop off the sampleprocessing component. Additional mechanisms known in the art includingbut not limited to adhesives, hook and loop fasteners, screws, or lockand groove configurations may be used.

In some embodiments, a component removal mechanism may be provided toassist with dropping off the sample processing component. Alternatively,no separate component removal mechanism may be required. In someinstances, a tip removal mechanism may be used as a component removalmechanism. In another example, a plunger may be used as a componentremoval mechanism. Alternatively, separate component removal mechanismsmay be provided. A component removal mechanism may use the principles ofgravity, friction, pressure, temperature, viscosity, magnetism, or anyother principles. A large quantity of tips can be stored within thedevice that are available as a shared resource to the pipette or robotto be utilized when required. Tips may be stored in a hopper, cartridge,or bandoleer to be used when required. Alternatively, tips may be storedin nested fashion to conserve space within the device. In anotherembodiment, a module can be configured to provide extra tips or anyother resources needed as a shared module in the device.

The fluid handling apparatus may interface with the sample processingcomponent at any number of interfaces. For example, the fluid handlingapparatus may interface with the sample processing component at one,two, three, four, five, six, seven, eight, nine, ten, or moreinterfaces. Each of the interfaces may be the same kind of interface, ormay be any combination of various interfaces (e.g., press fit, suction,magnetic, etc.). The number and/or type of interface may depend on thesample processing component. The fluid handling apparatus may beconfigured to interface with a sample processing component with one typeof interface, or may have multiple types of interface. The fluidhandling apparatus may be configured to pick up and/or transfer a singletype of sample processing component or may be capable of picking up andtransferring multiple types of sample processing components. The fluidhandling apparatus, assisted by the application of various tips, mayfacilitate or perform various sample processing tasks for or with thesample processing component, including physical and chemical processingsteps.

FIG. 52 provides an example of a fluid handling apparatus used to carrya sample processing component. The sample processing component may be acuvette carrier 5210. The cuvette carrier may have one or more interfacefeature 5212 that may be configured to interface with the fluid handlingdevice. In some embodiments, the interface feature may contact a pipettenozzle 5220 of the fluid handling device. A plurality of interfacefeatures may contact a plurality of pipette nozzles.

In some embodiments, a tip removal mechanism 5230 may be useful forremoving the cuvette carrier from the pipette nozzle. A plurality of tipremoval mechanisms may be actuated simultaneously or in sequence.

FIG. 53 shows a side view of a fluid handling apparatus useful forcarrying a sample processing component. A cuvette carrier 5310 mayinterface with the fluid handling apparatus. For example, nozzles 5320that may engage with the cuvette carrier. The nozzles may have the sameshape and/or configuration. Alternatively, the nozzles may have varyingconfigurations. The cuvette carrier may have one or more complementaryshape 5330, which may be configured to accept the nozzles. The nozzlesmay be engaged with the carrier through friction and/or vacuum assist.The nozzles may be for air displacement pipettes.

The cuvette carrier may interface with one or more cuvette 5340, orother types of vessels. The cuvette may have a configuration as shown inFIGS. 70A-B.

The fluid handling apparatus may also interface with a series ofconnected vessels. One such configuration is shown in FIG. 69, where thefluid handling apparatus may interface with pick-up ports 6920 to pickup the strip of vessels.

In some embodiments, a mini vessel is provided that may interface with apipette for various processing and analytical functions. The variousprocessing and analytics functions in some cases can be performed at apoint of service location.

Pick-Up Interface

A fluid handling device may be configured to interface with a tip or anyother component. As previously mentioned, a fluid handling device mayinclude a pipette nozzle, which may be press-fit to a pipette tip.Additional mechanisms may be used to connect a tip or other component tothe fluid handling device including, but not limited to, magnetic,snap-fit, hook and loop fasteners, elastics, ties, sliding mechanisms,locking mechanism, clamps, actuated mechanical components, and/oradhesives. The connection of a component or tip may permit the fluidhandling device to function as a robot capable of performing one or morefluid-handling or non-fluid handling functions. Such functions mayinclude the ability to transfer power to move tools or other objects,such as cartridges. When combined with a flexible supporting body (asdescribed above), the device may be able to perform such functionsacross a wide range of movement.

A pipette nozzle may be capable of interfacing with a single tip and/orvessel. For example, specific pipette nozzles may be configured tointerface with specific tips and/or vessels. Alternatively, a singlepipette nozzle may be capable of interfacing with a plurality of tipsand/or vessels. For example, the same pipette nozzle may be capable ofinterfacing with both a large and a small pipette tip and/or vessel. Apipette nozzle may be capable of interfacing with tips and/or vesselshaving different configurations, dimensions, volume capacities,materials, and/or size.

In one example, one or more rotational mechanism may be used. Suchrotational mechanisms may include screwing a tip onto a pipette nozzle.Such screwing mechanisms may employ external screws and/or internalscrews. FIG. 59 includes an example of a screw-mechanism. A pipettenozzle 5900 may be provided. A tip 5910 may be configured to connect tothe pipette nozzle. The tip may connect to the pipette nozzle directlyor via an interface 5920. In some embodiments, the interface may be anut or other connector. The interface 5920 may connect to the pipettenozzle 5900 in any manner including press-fit, screw, or any otherconnecting mechanism described elsewhere herein. Similarly, theinterface 5920 may connect to the tip 5910 via press-fit, screw, or anyother connecting mechanism described elsewhere herein.

In one example, a pipette tip 5910 may have an external screw ramp 5930.An interface 5920, such as a nut, may have a complementary internalscrew ramp 5940. In an alternate embodiment, the pipette tip may have aninternal screw ramp, and the interface, such as a nut, may have acomplementary external screw ramp. The pipette tip may be capable ofscrewing into an interior portion of the interface. A portion of anoutside surface of the pipette tip may contact an interior surface ofthe interface.

In an alternate embodiment, the pipette tip may be capable of screwingover an exterior portion of the interface. A portion of the insidesurface of the pipette tip may contact an exterior surface of theinterface. In such an embodiment, an interface may have an externalscrew ramp on its outer surface and/or an internal screw ramp on itsouter surface. The pipette tip may have a complementary internal screwramp on its internal surface or a complementary external screw ramp onits internal surface, respectively.

In additional embodiments, a portion of the tip surface may be embeddedin an interface, or a portion of the interface may be embedded withinthe tip.

A portion of the pipette nozzle may be within the interface, or aportion of the pipette nozzle may be external to the interface. In someembodiments, a portion of the pipette nozzle surface may be embeddedwithin a portion of the interface, or a portion of the interface surfacemay be embedded within a portion of the pipette nozzle.

A pipette nozzle 5900 may have one or more flanges 5950 or other surfacefeatures. Other examples of surface features may include grooves,protrusions, bumps, or channels. The flange may fit into a flange seatof a tip 5910. The flange may fit into the flange seat to preventrotation. This interface may be configured to prevent rotation of theinterface and tip once the tip is properly screwed in.

In alternate embodiments of the invention, no interface 5920 may berequired. A tip may screw directly into a pipette nozzle. The tip mayscrew directly over the nozzle, or inside the nozzle. An exteriorsurface of the tip may contact an interior surface of the nozzle, or aninternal surface of the tip may contact an external surface of thenozzle. In alternate embodiments, a portion of the tip surface may beembedded within a pipette nozzle, or a portion of a pipette nozzlesurface may be embedded within a tip.

A tip may have one, two or more external screw ramps. Any number ofexternal screw ramps may be provided. One, two, three, four, five, six,seven, eight, or more screw ramps may be provided. The screw ramps maybe external screw ramps, internal screw ramps, or any combinationthereof. The screw ramps may be equally radially spaced apart. A pipettetip may have one, two or more flange seats. One, two, three, four, five,six, seven, eight, or more flange seats may be provided. The flangeseats may be equally radially spaced apart. Alternatively, the intervalbetween flange seats may vary. The flange seats may be located radiallywhere a screw ramp reaches an end of a pipette tip. Alternatively, theflange seats may be located anywhere in relation to the screw ramps.

A pipette nozzle may have one, two or more flanges, or other surfacefeatures described elsewhere herein. One, two, three, four, five, six,seven, eight or more flanges may be provided. The flanges may be equallyradially spaced apart. Alternatively, the intervals between flanges mayvary. A flange may be configured to fit into a flange seat. In someembodiments, a one to one correspondence may be provided between flangesand flange seats. A first flange may fit into a first flange seat, and asecond flange may fit into a second flange seat. The flange seats mayhave complementary shapes to the flanges. In some embodiments, theflanges may have the same shape and the flange seats may fit over anyflange. Alternatively, the flanges may have different shapes and/orconfigurations so that specific flange seats may correspond to specificflanges.

In alternate embodiments, one or more flange may be provided within apipette nozzle. Complementary flange seats may be shaped on a pipettenozzle.

A flange may be press-fit into a flange seat. The connection between aflange and flange seat may be tight. Alternatively, a connection betweena flange and flange seat may be loose so that a flange may slide out ofa flange seat.

FIG. 60 provides an additional example of a nozzle-tip interfaceprovided in accordance with an embodiment of the invention. The pick-upand interface may use one or more features, characteristics, or methodsemployed within a ball-point pen-type configuration. A nozzle 6000 maybe configured to come into contact with a tip 6002. One or more pick-upclaw 6004 may be configured to pick up the tip. The pick-up claw mayhave one or more claw tine 6006 or other component that may grip or pickup the tip.

In some instances, a collar 6008 may fit over the pick-up claw 6004. Theclaw tines 6006 may extend out of the collar. The collar may have a clawcompression diameter 6010. The claw may slide within the pick-up collar.Thus, the tines may extend from the collar to varying amounts. The clawcompression diameter may compress the tines to come together. This mayenable the tines to grip an object, such as the tip, when the collarslides over the tines.

A ratchet mechanism 6012 may be provided. The ratchet mechanism mayslide over a portion of the claw. One or more claw pin 6014 may guidethe claw within the ratchet. For example, the claw pins may keep theclaw moving longitudinally along the ratchet, rather than slidingaround.

A claw spring 6016 may be provided, which may assist with providingforce along the claw in a longitudinal direction. In some instances, anozzle spring 6018 may be provided which may permit the nozzle to movein a longitudinal direction. The nozzle spring may optionally have asmaller diameter than the claw spring. The claw spring may wrap aroundthe outside a portion of the nozzle. One or more cap 6020 may beprovided.

A pick-up assembly, including the nozzle 6000, claw 6004, collar 6008,cap 6020 and associated portions may approach a tip 6002. The assemblymay press down to pick-up engage the tip. One or more tines 6006 of theclaw may capture a lip of the tip. The collar may be partially over thetines to compress the tines against the tip. The collar may slidefurther down to tighten the tines further around the tip in a pick-uppress step.

The assembly may then pull up. The tines may be caught on the lip of thetip in a pick-up lock step. The nozzle may force the tip against thetines, forming a seal. The entire assembly may be used in a pipettingfunction. For example, the pipette and connected tip may aspirate,dispense, and/or transfer a fluid. The claw may be locked in the collarduring the pipetting functions.

In order to remove the tip, the assembly may be pressed down in adrop-off engage step. In a drop-off pull away step, the assembly may belifted, with the collar sliding up relative to the claw, permitting thetines to loosen around the tip. The entire assembly may be lifted whilethe tip remains down, thereby separating the tip from the pick-upassembly.

FIG. 61 shows an example of an internal screw pick-up interface. A tip6100 may screw into a screw portion 6110 of the pipette. The portion maybe a pipette nozzle or interface between the tip and pipette nozzle. Thetip may include one or more flanges 6120 or other surface features. Anynumber or configuration of flanges may be provided, as describedelsewhere herein. The flanges may engage with one or more mechanism thatmay rotate the tip around a screw portion. Alternatively, the screwportion may spin while the tip remains stationary, optionally being heldin place using the flanges. The screw portion may include one or morescrews 6130 that may screw within the tip. Alternatively, the tip mayinclude one or more screws on its external surface and may screw intothe screw portion. The screw portion may include one or more fluidpathway 6140. The fluid pathway may be brought into fluid communicationwith the interior 6150 of the tip.

FIG. 62 illustrates an example of an O-ring tip pickup. A tip 6200 maybe picked up by a pipette nozzle 6210. A portion of the tip may fitwithin a portion of the nozzle. For example, a portion of the externalsurface of the tip may contact an internal surface of the nozzle.Alternatively, a portion of the nozzle may fit within a portion of thetip. For example, a portion of the internal surface of the tip maycontact an external surface of the nozzle.

The nozzle may have one or more O-ring 6220 that may contact the tip6200. The O-ring may be formed of an elastomeric material. The O-ringmay be provided around the circumference of the pipette nozzle.Alternatively, elastomeric material may be provided that need not beprovided around the entire circumference of the pipette nozzle. Forexample, one or more rubber balls or similar elastomeric protrusions maybe provided at one or more intervals within the pipette nozzle. Thepipette nozzle may have one or more groove into which one or moreO-rings may fit. Alternatively, the tip may have one or more grooves onits external surface into which one or more O-rings or other materialsmay fit.

A high-friction and/or flexible material may be provided between aportion of the nozzle and/or tip. This may enable the tip to bepress-fit into the nozzle, or for the nozzle to be press-fit into thetip. In some instances, both the nozzle and tip may have O-rings orsimilar materials. An O-ring may ensure a fluid seal between the tip andnozzle.

The pipette nozzle may have an internal shelf or flat back 6230. Theflat back may provide a physical stop to seat a tip in the appropriatelocation.

FIG. 63 provides an example of an expand/contract smart material tippickup. A tip 6300 may be picked up by a pipette nozzle 6310. A portionof the tip may fit within a portion of the nozzle. For example, aportion of the external surface of the tip may contact an internalsurface of the nozzle. Alternatively, a portion of the nozzle may fitwithin a portion of the tip. For example, a portion of the internalsurface of the tip may contact an external surface of the nozzle.

The nozzle may include a collar made of a magnetostrictive orelectrostrictive smart material which may contract when subject tomagnetic or electric field respectively. Electromagnetic coils, magneticfield manipulation, or a current generating power source may beincorporated to control the contraction and expansion of the material.

To pick up a tip, the nozzle may descent around the tip and the collarmay be activated, causing it to contract and grip the tip. The collarmay grip the tip tightly. The contraction of the collar may grip the tipsufficiently tightly to ensure a tight fluid seal. To release the tip,the collar may be deactivated to expand and release the tip.

The pipette nozzle may have an internal shelf or flat back 6320. Theflat back may provide a physical stop to seat a tip in the appropriatelocation.

In an alternate embodiment, the smart material of the nozzle may beinserted within a portion of the tip. The material may be activated tocause the material to expand and grip the tip from the inside. Thematerial may be deactivated to cause the material to contract andrelease the tip.

FIG. 64 provides an example of an expand/contract elastomer deflectiontip pickup. A tip 6400 may be picked up by a pipette nozzle 6410. Aportion of the tip may fit within a portion of the nozzle. For example,a portion of the external surface of the tip may contact an internalsurface of the nozzle. Alternatively, a portion of the nozzle may fitwithin a portion of the tip. For example, a portion of the internalsurface of the tip may contact an external surface of the nozzle.

The nozzle may include a rigid material 6420 and an elastomeric material6430. The rigid material may be a rigid block or solid material. The tipmay be surrounded by the elastomeric material. The rigid block may lieover the elastomeric material surrounding the tip.

An actuator may provide a force 6440 that may compress the rigid block6420. The rigid block may be pressed toward the tip. Pressing the rigidblock may compress the elastomer 6430, causing a bulging effect that mayshrink the internal chamber of the elastomer. Shrinking the internalchamber may cause the elastomer to securely grip the tip 6400.Compressing the elastomer in a first direction (e.g., toward the tip)may cause the elastomer to expand in a second direction (e.g.,perpendicular toward the tip), which may result in a compression of theelastomer around the tip.

In order to drop the tip off, the force 6440 may be removed, which maycause the rigid block to move away from the tip, and may release theelastomer from its compressed state.

FIG. 65 provides an example of a vacuum gripper tip pickup. A tip 6500may be provided, having a large head 6502. The large head may have alarge flat surface area.

The tip may engage with a nozzle 6510. The nozzle may have one or moretunnel 6520 therein. In some instances, one, two, three, four, five,six, seven, eight or more tunnels may be provided through the nozzle.The tunnels may be spaced radially equally apart, or at varyingintervals. The tunnels may have the same or differing diameters. A firstend of a tunnel may be coupled to a pressure source, while a second endof the tunnel may be facing the head 6502 of the tip. The pressuresource may be a negative pressure source. Tunnels may be connected to alower pressure region, creating a suction force, which may act on theflat head of the tip. The suction force may provide a pulling force thatmay act upwards to secure the tip to the nozzle.

In some embodiments, an O-ring 6530 may be provided. The O-ring or otherelastomeric member may be located between a nozzle and the head of atip. One or more groove or shelf may be provided in the nozzle and/ortip to accommodate the O-ring. The O-ring may permit a seal to be formedbetween the nozzle and tip. This may provide fluid tight seal between afluidic path 6540 within the nozzle and a fluid path 6550 within thetip.

In order to drop off the tip from the nozzle, the tunnels may bedisconnected from the negative suction pressure source. Alternatively,the pressure source itself may be turned off.

Such nozzle-tip connections and interfaces are provided by way ofexample only. Additional tip-nozzle interfaces, and/or variations orcombinations of those described herein may be implemented. In someembodiments, one or more components of a pipette may be configured to beexchangable. Such configurations may allow for future versions ofcomponents of a pipette (e.g. nozzles) with different functionality tobe added to or exchanged on the pipette.

Modular Fluid Handling

In some embodiments, one or more of the fluid handling apparatusconfigurations described elsewhere herein may be implemented in amodular fashion. For example, one or more pipette head may be providedin a modular format. In some embodiments, a single pipette module mayhave a single pipette head and/or nozzle thereon. Alternatively, asingle pipette module may have two, three, four, five, six or morepipette heads and/or nozzles thereon. Pipette modules may be stackednext to each other to form a multi-head configuration. Individualpipette modules may be removable, replaceable, and/or swappable.Individual pipette modules may each have the same configuration or mayhave different configurations. In some instances, different pipettemodules may be swapped out for others to provide differentfunctionality. Pipette modules described herein may also be referred toas “pipette cards,” “cards,” or “pipette units.”

FIG. 66 provides an example of a pipette module in accordance with anembodiment of the invention. The pipette module may include a pipettebody 6600 mounted on a support 6610. The support may include or moreguide rod 6612, track, screw, or similar feature. The pipette body maybe able to slide along the guide rod or similar feature. Any descriptionherein of guide rod may apply to any other feature that may guide themotion of a pipette body. In some instances, the pipette body may beable to travel upwards and/or downwards relative to the support alongthe guide rod

In some instances, the support may also include a lead screw 6614. Thelead screw may interact with an actuation interface 6602 of the pipettebody. The actuation interface may contact the lead screw, so that as thelead screw may turn, the actuation interface may engage with the teethof the screw and may cause the pipette body to move up or downcorrespondingly. In some embodiments, the actuation interface may be aspring-loaded flexure. The spring loaded flexure may be biased againstthe screw, thereby providing a strong flexible contact with the screw.The spring loaded flexture may be configured for precise kinematicconstraint. The screw may turn in response to an actuation mechanism. Insome embodiments, the actuation interface may be connected to thepipette piston by means of a magnet, offering sufficient degrees offreedom to limit wear and extend the life of the mechanism. In someembodiments, the actuation mechanism may be a motor, which may includeany type of motor described elsewhere herein. The motor may be directlyconnected to the screw or may be connected via a coupling. The actuationmechanism may move in response to one or more instructions from acontroller. The controller may be external to the pipette module, or maybe provided locally on the pipette module.

The pipette body 6600 may include a chassis. The chassis may optionallybe a shuttle clamshell chassis. A nozzle 6620 may be connected to thepipette body. The nozzle may extend from the pipette body. In someembodiments, the nozzle may extend downward from the pipette body. Thenozzle may have a fixed position relative to the pipette body.Alternatively, the nozzle may extend and/or retract from the pipettebody. The nozzle may have a fluid pathway therein. The fluid pathway maybe connected to a pipetting piston. Any descriptions of plungers,pressure sources, or fluid pathways described elsewhere herein may beused in a modular pipette. In some embodiments, the pipette body maysupport a motor 6630, geartrain, valve 6632, lead screw, magnetic pistonmounting block, piston cavity block and valve mount 6634, and/or othercomponents. One or more of the components described herein may beprovided within a chassis of the pipette body.

The pipette body may also include a guide rail 6640. The guide rail maypermit a portion of the pipette to move relative to the pipette body. Inone example, the pipette nozzle may move up or down relative to thepipette body. The pipette nozzle may be connected to an internalassembly that may move along the guide rail. In some embodiments, theguide rail 6640 may be configured to interface with another mechanismthat may prevent the pipette body from rotating. The guide rail may beconstrained by an exterior chassis, which may constrain rotation aboutthe guide rod.

FIG. 67A shows an example of modular pipette having a retracted shuttlein a full dispense position. A pipette body 6700 may be at an upwardposition relative to a support 6710. The pipette body may include anactuation interface 6702 that may engage with a lead screw 6714. When ashuttle is retracted, the actuation interface may be at the top of thelead screw. The mount may have a guide rod 6712 which may assist withguiding the pipette body relative to the mount.

FIG. 67B shows an example of modular pipette having a dropped shuttle ina full dispense position. A pipette body 6700 may be at a downwardposition relative to a support 6710. The pipette body may include anactuation interface 6702 that may engage with a lead screw 6714. When ashuttle is dropped, the actuation interface may be at the bottom of thelead screw. The mount may have a guide rod 6712 which may assist withguiding the pipette body relative to the mount.

The mount may be fully retracted, fully dropped, or have any positiontherebetween. The screw may turn to cause the pipette body to rise orlower relative to the mount. The screw may turn in a first direction tocause the pipette body to rise, and may turn in a second direction tocause the pipette body to drop. The screw may stop turning at any pointin order to provide a position of the pipette body. The pipette body maydrop with the nozzle, which may allow for greater complexity with lessrelative motion.

A plurality of pipette modules may be provided in a fluid handlingsystem. The pipette modules may have a blade configuration. A thin bladeform factor may be provided so that any number of blades may be stackedside by side in a modular fashion to create a pipetting system whereeach nozzle can work or move independently. A single blade may becomposed of multiple tools (nozzle, end effectors, etc.) that can bechosen for specific operations, thereby minimizing the space requiredfor the overall assembly. In some embodiments, a blade may also functionas a freezer, refrigerator, humidifier, and/or incubator for samplesand/or reagents held in vessels and/or cartridges.

The plurality of pipette modules may or may not be located adjacent toone another. In some embodiments, the pipette modules may be narrow andmay be stacked next to one another, to form a multi-head pipetteconfiguration. In some embodiments, a pipette module may have a width ofless than or equal to 1 μm, 5 μm, 10 μm, 50 μm, 100 μm, 300 μm, 500 μm,750 μm, 1 mm, 1.5 mm, 2 mm, 2.5 mm, 3 mm, 3.5 mm, 4 mm, 5 mm, 6 mm, 7mm, 8 mm, 9 mm, 1 cm, 1.5 cm, 2 cm, 3 cm, or 5 cm. Any number of pipettemodules may be positioned together. For example, one or more, two ormore, three or more, four or more, five or more, six or more, seven ormore, eight or more, nine or more, ten or more, twelve or more, fifteenor more, twenty or more, twenty-five or more, thirty or more, fifty ormore, seventy or more, one hundred or more pipette modules may bepositioned together. Additional pipette modules may be positionedseparately or together and optionally may have varying nozzles withdifferent dimensions and capabilities.

The separate pipette modules may be positioned adjacent to one anotherand may or may not contact one another. The pipette modules positionedtogether may or may not share a common support. The pipette bodies ofthe pipette modules may be able to move independently of one another upand down relative to the pipette mounts. The nozzles of the pipettemodules may be able to extend and/or retract independently relative tothe other pipette modules. In some embodiments, a pipette comprisingmultiple pipette modules on a common support may be configured such thatany one of the pipette modules is capable of contacting the samelocations within a device as may be contacted by one or more of theother pipette modules. This configuration may be desirable, for example,as a precaution for in the event that a pipette module becomesnon-functional, and it becomes desirable for another pipette module ofthe same common support to take over for the non-functional pipette.

The various pipette modules may have the same or differentconfigurations. The pipette nozzles of the pipette nozzles may be thesame or may vary. The pipette modules may be capable of interfacing withmultiple types of tips or with specialized tips. The pipette modules mayhave the same or varying degrees of sensitivity or coefficient ofvariation. The pipette modules may have the same or different mechanismsfor controlling the aspiration and/or dispensing of a fluid (e.g., airdisplacement, positive displacement, internal plunger, vertical plunger,horizontal plunger, pressure source). The pipette modules may have thesame or different mechanisms for picking up or removing a tip (e.g.,press-fit, screw-in, smart material, elastomeric material, click-fit, orany other interface described elsewhere herein or otherwise).

A modular pipette may have motion that may be broken down into aplurality of functions. For example motion may be broken into (1) motionof a piston and piston block in a (z) direction to aspirate and dispensefluid, and (2) motion of a shuttle assembly in a (z) direction to allowthe pipette module to engage with objects at various heights and provideclearance when moving in (xy) directions. In some embodiments, the (z)direction may be a vertical direction, and (xy) directions may behorizontal directions. The motion of the piston and piston block may beparallel to the motion of the shuttle assembly. Alternatively, themotion of the piston and piston block may be non-parallel and/orperpendicular. In other embodiments, the motion of the piston and pistonblock and/or the motion of the shuttle assembly may be horizontal or mayhave any other orientation.

Piston motion may be achieved in a very compact, flat package via theuse of a gear train and lead screw stacked horizontally, for example asillustrated in FIG. 66. A constant force spring, compression spring, orwave spring may be used to remove backlash in this assembly and maytherefore provide significantly improved accuracy/precision foraspiration and dispense. The system may use exact or very precisekinematic constraint with various springs in order to permit theassembly to operate precisely even with inaccuracies in the position orsize of each individual component.

All components which interact directly with the tips, nozzle, or pistonmay be mounted to a single “shuttle assembly” and this entire assemblymay move as one piece. The shuttle assembly may include a pipette body6600 as shown in FIG. 66. The various components may move with theshuttle assembly, which may be distinguishable from traditional pipetteswhere only the nozzle moves. This design may allow for simple, rigidconnection of these components to the critical piston/nozzle areawithout the need for complex linkages or relative motion between severalparts. It may also provide an expandable “platform” upon which tointegrate future components and functionalities.

The piston may be housed in a cavity. The cavity where the piston ishoused may be cut from a single piece of metal and any valves or nozzlesmay be mounted directly to this block. This may simplify the mounting ofcomponents that may be directly involved in the pipetting action and mayprovide a reliable air tight seal with little unused volume. This maycontribute to lower coefficients of variation for pipetting. Any of thecoefficient of variation values described elsewhere herein may beachieved by the pipette.

The shuttle assembly may be intentionally underconstrained in rotationabout a shuttle guide rod. This may assist with tolerating misalignmentin the device as the shuttle may have sufficient freedom to pivot sideto side (e.g., xy plane) into whatever position is needed to engage withtips or other interface objects.

The components in the shuttle assembly may be encased in a two piece“clamshell.” Some, more than half, or all of the components of theshuttle assembly may be encased within the clamshell. The clamshell caninclude two symmetric halves to the shuttle chassis that may hold thecomponents in place. It can also include a single half with deep pocketsfor component mounting and a flat second half that completes the processof securing components in place. The portions of the clamshell may ormay not be symmetric, or may or may not be the same thickness. Thesedesigns may allow the assembly to include a large number of smallcomponents without a complicated mounting method for each component. Theclamshell design may also allow for an assembly method where componentscan be simply dropped into their correct position and then the secondhalf of the clamshell may be put in place and fastened, thus lockingeverything in place. Additionally, this geometry lends itself to anapproach which integrates PCB routing boards directly into the clamshellchassis components in order to facilitate wiring for components insidethe device.

Any description of clamshell may apply to a multi-part housing or casingof the shuttle assembly. A housing of the shuttle assembly may be formedfrom one, two, three, four, five, six, seven, eight or more parts thatmay come together to form the housing. A clamshell may be an example ofa two-part shuttle housing. The portions of a clamshell may or may notbe connected by a hinge. The portions of the clamshell may be separablefrom one another.

In some embodiments, each nozzle/tip/piston/shuttle assembly may becombined into a single module (or blade) that is very thin and flat.This may allow stacking of several blades at a set distance from oneanother to create an arbitrarily large pipette. A desired number ofblades may be stacked together as needed, which may permit the pipetteto grow or shrink as needed. This modular approach can provide greatflexibility in the mechanical design since it breaks up functionalityand components into interchangeable parts. It may also enable modularcomponents in this design to be rapidly adapted for and integrated intonew pipettor systems; thus the same basic modular components can becapable of completing a large variety of tasks with differentrequirements. The modularization of functionality may also enable moreefficient device protocols due to fast and independent nozzle and pistoncontrol on board each pipette blade. This design may provide advantagesin servicing devices as defective blades can be swapped individually,rather than necessitating an entirely new pipettor. One or more of theblades may be independently movable and/or removable relative to theother places.

FIG. 67C shows yet another embodiment wherein a plurality of individualpipette units 6720 are provided. FIG. 67C is a front view showing thateach of the individual pipette units 6720 may be individually movablerelative to any other pipette unit in the pipette chassis 6722. Some ofthe individual pipette units 6724 are configured to be larger volumeunits and use larger head units 6726. Each of the pipette units 6720 and6724 can be moved up and down individually as indicated by arrow 6728.The system may optionally have imaging devices 6730 and 6732 to viewactivity at the pipette tips. This can be used as quality control toimage whether a tip is properly seated on the pipette nozzle, whethersufficient volume of sample is in the tip, whether there is undesiredbubbles or other defects in the samples. In the present embodiment, theplurality of imaging devices 6730 and 6732 are sufficient to image allof the tips of the pipette nozzle.

FIG. 67D shows a side view of one embodiment an individual pipette unit6720. FIG. 67D shows that this pipette unit 6720 may have aforce-providing unit 6740 such as but not limited to a motor, apiezoelectric drive unit, or the like. Although direct drive is notexcluded, the present embodiment uses a transmission such as but notlimited to pulleys, linkages, or gears 6744 and 6746 are used to turn alead screw 6748 that in turn moves the piston slide mechanism 6750 whichcan move up and down as indicated by arrow 6752. This in turn moves apiston 6754 that drives, using direct or air displacement, theaspiration or dispensing of fluid in tips (not shown) coupled to thenozzle portion 6756. A tip ejector slide 6760 is actuated when the lowerextending portion of the piston slide mechanism 6750 pushes down on andmoves the tip ejector slide 6760 down as indicated by arrow 6762. Afterthe tip is ejected, the slide 6760 may return to its original position.

As indicated by arrow 6770, the entire pipette unit 6720 can translateup and down in a first frame of reference. Components within the pipetteunit 6720 can also move up and down in a second frame of reference. Theaspirating and dispensing of liquid is independent of the movement ofthe unit 6720. The present embodiment also shows that there is no tubingextending to an external source. All fluid is kept separate from theinternals of the pipette unit 6720 so that the units can be used withouthaving to be cleaned or washed between uses. Some embodiments may havehydrophobic coatings, seals, filters, filter paper, frits, septa, orother fluid sealing items to prevent fluid and aerosolized particlesfrom entering the hardware, non-disposable portions of the pipettes.

In some embodiments, fully modular pipette unit 6720 for various fluidvolumes and tip types can be provided with a common drive train design.In one embodiment, nozzle and all fluid components (including thepiston/pump) are all located in a self-contained module which can bebuilt and validated outside the rest of the assembly. The commonplatform allows for future versions of nozzles with differentfunctionality to be added to the system through either new tips that canengage the heads or by replacing the module pipette unit 6720 with anupdated pipette unit, so long as the interfaces both mechanical andelectrical remain compatible with what is on the pipette chassis.

Pipette units may be optimized to pipette different volumes of fluid.Pipette units may have different volume capacities. In some embodiments,the volume capacity of a pipette unit is related to the volume of thepiston block or piston, the nozzle of the pipette unit, and/or tipswhich interface with the nozzle. In some embodiments, a pipette unit mayhave a pipetting capacity as low as 0.1 microliter or as high as 20mililiters, or any volume between. In some embodiments, a pipette unitmay be optimized for pipetting a range of volumes, including, forexample, 0.1-2 microliters; 0.1-10 microliters; 1-10 microliters; 1-50microliters; 2-20 microliters; 1-100 microliters; 10-200 microliters;20-200 microliters; or 100-1000 microliters.

Sensor Probes

In some embodiments, a pipette, pipette unit or any other component of adevice described herein may contain a probe. The probe may include oneor more sensors, e.g. for motion, pressure, temperature, images, etc.Integration of a probe into one or more components of a device may aidin monitoring one or more conditions or events within a device. Forexample, a touch probe may be integrated with a pipette, such that whena pipette is moved it may sense its location (e.g. through pressure,motion, or imaging). This may increase the precision and accuracy andlower the COV of movement of the pipette. In another example, a probe ona pipette may obtain information regarding the strength of a sealbetween a pipette nozzle and a pipette tip. In another example, a probemay contain a temperature sensor. If the probe is attached, for example,to a centrifuge, cartridge, or pipette, the probe may obtain informationregarding the temperature of the area in the vicinity of the centrifuge,cartridge, or pipette. A probe may be in communication with a controllerof a module, device, or system, such that information obtained by theprobe may be sent to the controller. The controller may use thisinformation in order to calibrate or optimize device performance. Forexample, if a probe senses that a tip is not properly sealed on apipette nozzle, the controller may direct the tip to be ejected from thepipette nozzle, and for a new pipette tip to be loaded onto the nozzle.In some embodiments, a probe may have a stand-alone structure, and notbe integrated with another component of a device.

Vessels/Tips

A system may comprise one, two or more vessels and/or tips, or maycontain a device that may comprise one, two or more vessels and/or tips.One or more module of a device may comprise one, two or more vesselsand/or tips.

A vessel may have an interior surface and an exterior surface. A vesselmay have a first end and a second end. In some embodiments, the firstend and second ends may be opposing one another. The first end or secondend may be open. In some embodiments, a vessel may have an open firstend and a closed second end. In some embodiments, the vessel may haveone or more additional ends or protruding portions which may be open orclosed. In some embodiments, a vessel may be used to contain a substratefor an assay or reaction. In other embodiments, the substrate itself mayfunction as a sort of vessel, obviating the need for a separate vessel.

The vessel may have any cross-sectional shape. For example, the vesselmay have a circular cross-sectional shape, elliptical cross-sectionalshape, triangular cross-sectional shape, square cross-sectional shape,rectangular cross-sectional shape, trapezoidal cross-sectional shape,pentagonal cross-sectional shape, hexagonal cross-sectional shape, oroctagonal cross-sectional shape. The cross-sectional shape may remainthe same throughout the length of the vessel, or may vary.

The vessel may have any cross-sectional dimension (e.g., diameter,width, or length). For example, the cross-sectional dimension may beless than or equal to about 0.1 mm, 0.5 mm, 1 mm, 1.5 mm, 2 mm, 2.5 mm,3 mm, 3.5 mm, 4 mm, 4.5 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 1 cm, 1.2 cm,1.5 cm, 2 cm, or 3 cm. The cross-sectional dimension may refer to aninner dimension or an outer dimension of the vessel. The cross-sectionaldimension may remain the same throughout the length of the vessel or mayvary. For example, an open first end may have a greater cross-sectionaldimension than a closed second end, or vice versa.

The vessel may have any height (wherein height may be a dimension in adirection orthogonal to a cross-sectional dimension). For example, theheight may be less than or equal to about 0.1 mm, 0.5 mm, 1 mm, 1.5 mm,2 mm, 2.5 mm, 3 mm, 3.5 mm, 4 mm, 4.5 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm,1 cm, 1.2 cm, 1.5 cm, 2 cm, 3 cm, 4 cm, 5 cm, 6 cm, 7 cm, 8 cm, 9 cm, or10 cm. In some embodiments, the height may be measured between the firstand second ends of the vessel.

The interior of the vessel may have a volume of about 1,000 μL or less,500 μL or less, 250 μL or less, 200 μL or less, 175 μL or less, 150 μLor less, 100 μL or less, 80 μL or less, 70 μL or less, 60 μL or less, 50μL or less, 30 μL or less, 20 μL or less, 15 μL or less, 10 μL or less,8 μL or less, 5 μL or less, 1 μL or less, 500 nL or less, 300 nL orless, 100 nL or less, 50 nL or less, 10 nL or less, 1 nL or less, 500 pLor less, 250 pL or less, 100 pL or less, 50 pL or less, 10 pL or less, 5pL or less, or 1 pL or less.

One or more walls of the vessel may have the same thickness or varyingthicknesses along the height of the vessel. In some instances, thethickness of the wall may be less than, and/or equal to about 1 μm, 3μm, 5 μm, 10 μm, 20 μm, 30 μm, 50 μm, 75 μm, 100 μm, 200 μm, 300 μm, 400μm, 500 μm, 600 μm, 700 μm, 800 μm, 900 μm, 1 mm, 1.5 mm, 2 mm, or 3 mm.

One or more vessels may be provided which may have the same shape and/orsize, or varying shapes and/or sizes.

A vessel may be formed of a single integral piece. Alternatively, thevessel may be formed from two or more vessel pieces. The two or morevessel pieces may be permanently attached to one another, or may beselectively separable from one another. A vessel may include a body anda cap. Alternatively, some vessels may only include a body.

A vessel may be configured to contain and/or confine a sample. A vesselmay be configured to engage with a fluid handling system. Any fluidhandling system known in the art, such as a pipette, or embodimentsdescribed elsewhere herein may be used. In some embodiments, a vesselmay be configured to engage with a tip that may be connected to a fluidhandling device, such as a pipette. A vessel may be configured to acceptat least a portion of a tip within the vessel interior. A tip may beinserted at least partway into the vessel. In some embodiments, the tipmay be configured to enter the vessel all the way to the bottom of thevessel. Alternatively, the tip may be configured to be inserted no morethan part way into the vessel.

Vessel material can be of different types, depending on the propertiesrequired by the respective processes. Materials may include but notlimited to: polymers, semiconductor materials, metals, organicmolecules, ceramics, composites, laminates, etc. The material may berigid or flexible, or able to transition between the two. Vesselmaterials may include, but not limited to polystyrene, polycarbonate,glass, metal, acrylics, semiconductor materials, etc., and may includeone of several types of coatings. Vessel materials may be permeable toselective species by introducing functionalized pores on the vesselwalls. These allow certain molecular species to pass through thematerial. Vessel material can also be coated to prevent absorption ofsubstances such as water. Other coatings might be used to achievespecific optical characteristics such as transmission, reflectance,fluorescence, etc.

Vessel can be of different geometries including, but not limited to,rectangular, cylindrical, hexagonal, and may include, withoutlimitation, attributes such as perforations, permeable membranes,particulates or gels depending on the application. Vessels may becomprised of microfluidic channels or electrical circuits, optionally ona silicon substrate.

Vessels may also be active and perform a set of tasks. Vessels maycontain active transporters to pump fluids/suspensions throughmembrane/septal barriers.

Vessels may be designed to have specific opticalproperties—transparency, opacity, fluorescence, or other propertiesrelated to any part of the electromagnetic spectrum. Vessels may bedesigned to act as locally heated reactors by designing the material toabsorb strongly in the infrared part of the electromagnetic spectrum.

Vessel walls might be designed to respond to different electromagneticradiation—either by absorption, scattering, interference, etc.Combination of optical characteristics and embedded sensors can resultin vessels being able to act as self-contained analyzers—e.g.,photosensitive material on vessel walls, with embedded sensors willtransform a vessel into a spectrophotometer, capable of measuringchanges in optical signals.

In some embodiments, vessels can be thought of as intelligent containerswhich can change their properties by “sampling” the surrounding fluids.Vessels could allow for preferential ion transfer between units, similarto cells, signaled by electrical and/or chemical triggers. They couldalso influence containment of the fluid inside it in response toexternal and/or internal stimuli. Response to stimuli may also result inchange of size/shape of the vessel. Vessels might be adaptive inresponse to external or internal stimuli, and might enable reflextesting by modification of assay dynamic range, signal strength, etc.

Vessels can also be embedded with different sensors or have differentsensors embedded in them, such as environmental (temperature, humidity,etc.), optical, acoustic, or electro-magnetic sensors. Vessels can bemounted with tiny wireless cameras to instantly transmit informationregarding its contents, or alternatively, a process which happens in it.Alternatively, the vessel can comprise another type of detector ordetectors, which transmit data wirelessly to a central processing unit.

Vessels can be designed for a range of different volumes ranging from afew microliters to milliliters. Handling fluids across different lengthand time scales involves manipulating and/or utilizing variousforces—hydrodynamic, inertial, gravity, surface tension,electromagnetic, etc. Vessels may be designed to exploit certain forcesas opposed to others in order to manipulate fluids in a specific way.Examples include use surface tension forces in capillaries to transferfluids. Operations such as mixing and separation require differentstrategies depending on volume—vessels may be designed to specificallytake advantage of certain forces. Mixing, in particular is importantwhile handling small volumes, since inertial forces are absent. Novelmixing strategies such as using magnetic particles with externalforcing, shear-induced mixing, etc. might be adopted to achieveefficient mixing.

Vessels offer flexibility over microfluidic chips due to their inherentflexibility in handling both small and large volumes of fluids.Intelligent design of these vessels allows us to handle a larger rangeof volumes/sizes compared to microfluidic devices. In one embodiment,vessels were designed with tapered bottoms. This taper is in at leastthe interior surface of the vessel. It should be understood that theexterior may be tapered, squared, or otherwise shaped so long as theinterior is tapered. These features reduce sample/liquid overages thatare needed. Namely, small volumes can be mixed in the vessel andextracted without wasting/leaving behind residual liquid. This designallows one to work with both small volumes and larger volumes ofliquids. In addition, vessels can take advantage of forces whichmicrofluidic devices cannot—thereby offering more flexibility inprocessing. Vessels may also offer the ability to dynamically changescales, by switching to different sizes. In the “smart vessel” concept,the same vessel can change capacity and other physical attributes totake advantage of different forces for processing fluids. This actuationcan be programmed, and externally actuated, or initiated by changes influid inside.

The functionality of a vessel can go beyond fluid containment—differentvessels can communicate via surface features or external actuation andengage in transport of fluids/species across vessel boundaries. Thevessel thus becomes a vehicle for fluid containment, processing, andtransport—similar to cells. Vessels can fuse in response to externalactuation and/or changes in internal fluid composition. In thisembodiment, vessels can be viewed as functional units, capable ofexecuting on or several specialized function—separations such asisoelectric focusing, dialysis, etc. Vessels can be used to samplecertain fluids and generate information regarding transformations, endpoints, etc.

Vessels can act as self-contained analytical units, with in-builtdetectors and information exchange mechanisms, through sensors andtransmitters embedded inside vessel walls. Vessel walls can be made withtraditional and/or organic semiconductor materials. Vessels can beintegrated with other sensors/actuators, and interface with othervessels. A vessel, in this embodiment, can be viewed as a system capableof containment, processing, measurement, and communication. In someembodiments, a vessel may contain a chip for electric manipulation ofvery small volumes of liquid.

Vessels can also have sample extraction, collection, and fluid transferfunctionalities. In this embodiment, a vessel would act like a pipettebeing stored in the cartridge, and able to transfer fluid to a specificlocation. Examples include a viral transport medium for nucleic acidamplification assays, where the vessel is used to both collect andtransport the viral transport medium. Another example would be a cuvettecoming out of the device in order to collect a fingerstick sample.

Vessels may be designed to contain/process various sample typesincluding, but not limited to blood, urine, feces, etc. Different sampletypes might require changes in vessel characteristics—materials, shape,size, etc. In some embodiments, vessels perform sample collection,processing, and analysis of contained sample.

A vessel or subvessel may be sealed with or otherwise contain reagentsinside it. A pipette may act to release the reagent from the vessel whenneeded for a chemical reaction or other process, such as by breaking theseal that contains the reagent. The vessels may be composed of glass orother material. A reagent that would otherwise be absorbed intotraditional polymer tips or degrade when exposed to the environment maynecessitate such compartmentalization or sealing in a vessel.

In some embodiments, vessels provided herein may have rounded edges tominimize fluid loss during fluid handling.

A vessel (e.g. a tip) may have an interior surface and an exteriorsurface. A vessel (e.g. a tip) may have a first end and a second end. Insome embodiments, the first end and the second ends may be opposing oneanother. The first end and/or second end may be open. A vessel (e.g. atip) may include a passageway connecting the first and second ends. Insome embodiments, a vessel (e.g. a tip) may include one or moreadditional ends or protrusions. For example, the vessel (e.g. a tip) mayhave a third end, fourth end, or fifth end. In some embodiments, the oneor more additional ends may be open or closed, or any combinationthereof.

The vessel (e.g. a tip) may have any cross-sectional shape. For example,the vessel may have a circular cross-sectional shape, ellipticalcross-sectional shape, triangular cross-sectional shape, squarecross-sectional shape, rectangular cross-sectional shape, trapezoidalcross-sectional shape, pentagonal cross-sectional shape, hexagonalcross-sectional shape, or octagonal cross-sectional shape. Thecross-sectional shape may remain the same throughout the length of thevessel (e.g. a tip), or may vary.

The vessel (e.g. a tip) may have any cross-sectional dimension (e.g.,diameter, width, or length). For example, the cross-sectional dimensionmay be less than or equal to about 0.1 mm, 0.5 mm, 1 mm, 1.5 mm, 2 mm,2.5 mm, 3 mm, 3.5 mm, 4 mm, 4.5 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 1 cm,1.2 cm, 1.5 cm, 2 cm, or 3 cm. The cross-sectional dimension may referto an inner dimension or an outer dimension of the vessel (e.g. a tip).The cross-sectional dimension may remain the same throughout the lengthof the vessel (e.g. a tip) or may vary. For example, an open first endmay have a greater cross-sectional dimension than an open second end, orvice versa. The cross-sectional dimension ratio of the first end to thesecond end may be less than, and/or equal to about 100:1, 50:1, 20:1,10:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:10, 1:20, 1:50 or1:100. In some embodiments, the change in the cross-sectional dimensionmay vary at different rates.

The vessel (e.g. a tip) may have any height (wherein height may be adimension in a direction orthogonal to a cross-sectional dimension). Forexample, the height may be less than, or equal to about 0.1 mm, 0.5 mm,1 mm, 1.5 mm, 2 mm, 2.5 mm, 3 mm, 3.5 mm, 4 mm, 4.5 mm, 5 mm, 6 mm, 7mm, 8 mm, 9 mm, 1 cm, 1.2 cm, 1.5 cm, 2 cm, 3 cm, 4 cm, 5 cm, 6 cm, 7cm, 8 cm, 9 cm, or 10 cm. In some embodiments, the height may bemeasured between the first and second ends of the tip.

The interior of the vessel (e.g. a tip) may have a volume of about 1,000μL or less, 500 μL or less, 250 μL or less, 200 μL or less, 175 μL orless, 150 μL or less, 100 μL or less, 80 μL or less, 70 μL or less, 60μL or less, 50 μL or less, 30 μL or less, 20 μL or less, 15 μL or less,10 μL or less, 8 μL or less, 5 μL or less, 1 μL or less, 500 nL or less,300 nL or less, 100 nL or less, 50 nL or less, 10 nL or less, 1 nL orless, 500 pL or less, 250 pL or less, 100 pL or less, 50 pL or less, 10pL or less, 5 pL or less, or 1 pL or less.

One or more walls of the vessel (e.g. a tip) may have the same thicknessor varying thicknesses along the height of the vessel (e.g. a tip). Insome instances, the thickness of the wall may be less than and/or equalto about 1 μm, 3 μm, 5 μm, 10 μm, 20 μm, 30 μm, 50 μm, 75 μm, 100 μm,200 μm, 300 μm, 400 μm, 500 μm, 600 μm, 700 μm, 800 μm, 900 μm, 1 mm,1.5 mm, 2 mm, or 3 mm.

One or more vessels (e.g. a tip) may be provided which may have the sameshape and/or size, or varying shapes and/or sizes. Any of the variousembodiments described herein may have one or more features of thevessels and/or tips as described elsewhere herein.

A tip may be formed of a single integral piece. Alternatively, the tipmay be formed from two or more tip pieces. The two or more tip piecesmay be permanently attached to one another, or may be selectivelyseparable from one another. Chemistries or sensors may also bephysically integrated into a tip, effectively enabling a completelaboratory test on a vessel (e.g. a tip). Vessels (e.g. a tip) may eachindividually serve different preparatory, assay, or detection functions.Vessels (e.g. a tip) may serve multiple functions or all functionswithin a single vessel or tip.

A vessel (e.g. a tip) may be formed of a material that may be rigid,semi-rigid, or flexible. The vessel (e.g. a tip) may be formed ofmaterial that is conductive, insulating, or that incorporates embeddedmaterials/chemicals/etc. The vessel (e.g. a tip) may be formed of thesame material or of different materials. In some embodiments, the vessel(e.g. a tip) may be formed of a transparent, translucent, or opaquematerial. The inside surface of a tip can be coated with reactants thatare released into fluids; such reactants can be plated, lyopholized,etc. The vessel (e.g. a tip) may be formed of a material that may permita detection unit to detect one or more signals relating to a sample orother fluid within the vessel (e.g. a tip). For example, the vessel(e.g. a tip) may be formed of a material that may permit one or moreelectromagnetic wavelength to pass therethrough. Examples of suchelectromagnetic wavelengths may include visible light, IR, far-IR, UV,or any other wavelength along the electromagnetic spectrum. The materialmay permit a selected wavelength or range(s) of wavelengths to passthrough. Examples of wavelengths are provided elsewhere herein. Thevessel (e.g. a tip) may be transparent to permit optical detection ofthe sample or other fluid contained therein.

The vessel (e.g. a tip) may form a wave guide. The vessel (e.g. a tip)may permit light to pass through perpendicularly. The vessel (e.g. atip) may permit light to pass through along the length of the vessel.The vessel (e.g. a tip) may permit light to light to enter and/or travelat any angle. In some embodiments, the vessel (e.g. a tip) may permitlight to enter and/or travel at selected angles or ranges of angles. Thevessel and/or tip may form one or more optic that may focus, collimate,and/or disperse light.

The material may be selected to be impermeable to one or more fluids.For example, the material may be impermeable to the sample, and/orreagents. The material may be selectively permeable. For example, thematerial may permit the passage of air or other selected fluids.

Examples of materials used to form the vessel and/or tip may includefunctionalized glass, Si, Ge, GaAs, GaP, SiO₂, SiN₄, modified silicon,or any one of a wide variety of gels or polymers such as(poly)tetrafluoroethylene, (poly)vinylidenedifluoride, polystyrene,polycarbonate, polypropylene, polymethylmethacrylate (PMMA), ABS, orcombinations thereof. In an embodiment, an assay unit may comprisepolystyrene. The materials may include any form of plastic, or acrylic.The materials may be silicon-based. Other appropriate materials may beused in accordance with the present invention. Any of the materialsdescribed here, such as those applying to tips and/or vessels may beused to form an assay unit. A transparent reaction site may beadvantageous. In addition, in the case where there is an opticallytransmissive window permitting light to reach an optical detector, thesurface may be advantageously opaque and/or preferentially lightscattering.

Vessels and/or tips may have the ability to sense the liquid leveltherein. For example, vessels and/or tips may have capacitive sensors orpressure gauges. The vessels may employ any other technique known in theart for detecting a fluid level within a container. The vessels and/ortips may be able to sense the liquid level to a high degree ofprecision. For example, the vessel and/or tip may be able to detect aliquid level to within about 1 nm, 5 nm, 10 nm, 50 nm, 100 nm, 150 nm,300 nm, 500 nm, 750 nm, 1 μm, 3 μm, 5 μm, 10 μm, 50 μm, 75 μm, 100 μm,150 μm, 200 μm, 250 μm, 300 μm, 400 μm, 500 μm, 600 μm, 700 μm, 800 μm,900 μm, or 1 mm.

A tip may assist with the dispensing and/or aspiration of a sample. Atip may be configured to selectively contain and/or confine a sample. Atip may be configured to engage with a fluid handling device. Any fluidhandling system known in the art, such as a pipette, or embodimentsdescribed elsewhere herein may be used. The tip may be connected to thefluid handling device to form a fluid-tight seal. In some embodiments,the tip may be inserted into a vessel. The tip may be inserted at leastpartway into the vessel. The tip may include a surface shape or featurethat may determine how far the tip can be inserted into the vessel.

Vessels and/or tips may be independently formed and may be separate fromone another. Vessels and/or tips may be independently movable relativeto one another. Alternatively, two or more vessels and/or tips may beconnected to one another. They may share a common support. For example,the two or more vessels and/or tips may be cut from a samematerial—e.g., cut into a common substrate. In another example, two ormore vessels and/or tips may be directly linked adjacent to one anotherso that they directly contact one another. In another example, one ormore linking component may link the two or more vessels and/or tipstogether. Examples of linking components may include bars, strips,chains, loops, springs, sheets, or blocks. Linked vessels and/or tipsmay form a strip, array, curve, circle, honeycombs, staggered rows, orany other configuration. The vessels and/or connections may be formed ofan optically transparent, translucent, and/or opaque material. In someinstances, the material may prevent light from entering a space withinthe vessels and/or cavities. Any discussion herein of vessels and/ortips may apply to cuvettes and vice versa. Cuvettes may be a type ofvessel.

FIG. 69 provides an example of a vessel strip. The vessel strip providesan example of a plurality of vessels that may be commonly linked. Thevessel strip 6900 may have one or more cavities 6910. The cavities mayaccept a sample, fluid or other substance directly therein, or mayaccept a vessel and/or tip that may be configured to confine or accept asample, fluid, or other substance therein. The cavities may form a row,array, or any other arrangement as described elsewhere herein. Thecavities may be connected to one another via the vessel strip body.

The vessel strip may include one or more pick-up interface 6920. Thepick-up interface may engage with a sample handling apparatus, such as afluid handling apparatus. The pick-up interface may interface with oneor more pipette nozzle. Any of the interface configurations describedelsewhere herein may be used. For example, a pipette nozzle may bepress-fit into the pick-up interface. Alternatively, the pick-upinterface may interface with one or more other component of the pipette.

The vessel strip may be useful for colorimetric analysis or cytometry.The vessel strip may be useful for any other analysis describedelsewhere herein.

FIGS. 70A and 70B provide another example of a cuvette 7000. The cuvetteprovides an example of a plurality of channels that may be commonlylinked. The cuvette carrier may have a body formed from one, two or morepieces. In one example, a cuvette may have a top body portion 7002 a,and a bottom body portion 7002 b. The top body portion may have one ormore surface feature thereon, such as a cavity, channel, groove,passageway, hole, depression, or any other surface feature. The bottombody portion need not include any surface features. The bottom bodyportion may be a solid portion without cavities. The top and bottom bodyportion may come together to form a cuvette body. The top and bottombody portion may have the same footprint, or may have differingfootprints. In some instances, the top body portion may be thicker thanthe bottom body portion. Alternatively, the bottom body portion may bethicker or equal in thickness to the top body portion.

The cuvette 7000 may have one or more cavities 7004. The cavities mayaccept a sample, fluid or other substance directly therein. The cavitiesmay form a row, array, or any other arrangement as described elsewhereherein. The cavities may be connected to one another via the cuvettebody. In some instances, the bottom of a cavity may be formed by abottom body portion 7002 b. The walls of a cavity may be formed by a topbody portion 7002 a.

The cuvette may also include one or more fluidically connected cavities7006. The cavities may accept a sample, fluid or other substancedirectly therein, or may accept a vessel and/or tip (e.g., cuvette) thatmay be configured to confine or accept a sample, fluid, or othersubstance therein. The cavities may form a row, array, or any otherarrangement as described elsewhere herein. The cavities may befluidically connected to one another via a passageway 7008 through thecuvette body.

The passageway 7008 may connect two cavities, three cavities, fourcavities, five cavities, six cavities, seven cavities, eight cavities,or more. In some embodiments, a plurality of passageways may beprovided. In some instances, a portion of the passageway may be formedby a top body portion 7002 a, and a portion of the passageway may beformed by a bottom body portion 7002 b. The passageway may be orientedin a direction that is not parallel (e.g., is parallel) to anorientation of a cavity 7006 to which it connects. For example, thepassageway may be horizontally oriented while a cavity may be verticallyoriented. The passageway may optionally permit a fluid to flow from onefluidically connected cavity to another.

The cuvette may include one or more pick-up interface. Optionally, apick-up interface may be one or more cavity, 7004, 7006 of the cuvette.The pick-up interface may engage with a sample handling apparatus, suchas a fluid handling apparatus. The pick-up interface may interface withone or more pipette nozzle. Any of the interface configurationsdescribed elsewhere herein may be used. For example, a pipette nozzlemay be press-fit into the pick-up interface, or the nozzle may interactmagnetically with the pick-up interface. Alternatively, the pick-upinterface may interface with one or more other component of the pipette.Optionally, the cuvette may include embedded magnet(s) or magneticfeature(s) that allow for a sample handling apparatus to pickup and/ordropoff the cuvette based on magnetic forces. In some embodiments, asample handling apparatus may directly transfer a cuvette from acartridge to a cytometry station. In some embodiments, a module-levelsample handling system may transfer a cuvette from an assay station to acytometry station or detection station in the same module. In someembodiments, a device-level sample handling system may transfer acuvette from an assay station to a cytometry station or detectionstation in a different module.

Cuvettes may be useful, for example, for colorimetric analysis orcytometry. The cuvette may be useful for any other analysis describedelsewhere herein. In some embodiments, a cuvette has a configurationoptimized for use with a cytometer, e.g. to interface with a microscopystage. In some embodiments, a cuvette has a configuration optimized foruse with a spectrophotometer.

A cuvette may be formed of any material, including those describedelsewhere herein. The cuvette may optionally be formed of a transparent,translucent, opaque material, or any combination thereof. The cuvettemay prevent a chemical contained therein from passing from one cavity toanother.

Referring now to FIG. 92, a still further embodiment of a cuvette willnow be described. FIG. 92 shows a cuvette 7030 with a plurality ofreaction wells 7032. It further includes side walls that allow foroptical, colorimetric, turbidimetric, or other visual observation, andoptionally, non-visual sensing of sample therein. In the presentembodiment, the cuvette 7030 has at least one elevated portion 7040 thatallows for engagement with a transport mechanism such as a pipette tomove the cuvette from one location to another. The elevated portion 7040allows the portion of the cuvette 7030 with the reaction vessels/wellsto be positioned lower in the detector, facilitating detector design andmore shielding the sample from outside light or other undesired externalconditions during measurement. In some embodiments, the cuvette 7030 mayhave ledges, legs, or other stability features on the top and/or bottomportions so that it can support itself against a bottom surface or sidewall surface of the detector station if the pipette or other transportmechanism disengages it so that the pipette or other mechanism canperform other tasks such as but not limited to pipetting or transportingother samples or reagents.

Referring now to FIG. 93, this embodiment shows that the lift location7040 of the cuvette is centrally located to provide a more balancedcondition when using only a single nozzle of the fluid transport systemto move the cuvette 7038.

FIG. 71 shows an example of a tip in accordance with an embodiment ofthe invention. The tip 7100 may be capable of interfacing with amicrocard, cuvette carrier and/or strip, including any examplesdescribed herein.

The tip may include a narrow portion that may deposit a sample 7102, asample volume area 7104, and/or a nozzle insertion area 7106. In someinstances, the tip may include one or more of the areas described. Thesample deposit area may have a smaller diameter than a sample volumearea. The sample volume area may have a smaller volume than a nozzleinsertion area. The sample deposit area may have a smaller volume than anozzle insertion area.

In some embodiments, a lip 7108 or surface may be provided at an end ofthe nozzle insertion area 7106. The lip may protrude from the surface ofthe nozzle insertion area.

The tip may include one or more connecting region, such as a funnelregion 7110 or step region 7112 that may be provided between varioustypes of area. For example, a funnel region may be provided between asample deposit area 7102 and a sample volume area 7104. A step region7112 may be provided between a sample volume area 7104, and a nozzleinsertion area. Any type of connecting region may or may not be providedbetween the connecting regions.

A sample deposit area may include an opening through which a fluid maybe aspirated and/or dispensed. A nozzle insertion area may include anopening into which a pipette nozzle may optionally be inserted. Any typeof nozzle-tip interface as described elsewhere herein may be used. Theopening of the nozzle insertion area may have a greater diameter than anopening of the sample deposit area.

The tip may be formed of a transparent, translucent, and/or opaquematerial. The tip may be formed from a rigid or semi-rigid material. Thetip may be formed from any material described elsewhere herein. The tipmay or may not be coated with one or more reagents.

The tip may be used for nucleic acid tests, or any other tests, assays,and/or processes described elsewhere herein.

FIG. 72 provides an example of a test strip. The test strip may includea test strip body 7200. The test strip body may be formed from a solidmaterial or may be formed from a hollow shell, or any otherconfiguration.

The test strip may include one or more cavities 7210. In someembodiments, the cavities may be provided as a row in the body. Thecavities may optionally be provided in a straight row, in an array(e.g., m×n array where m, n are whole numbers greater than zeroincluding but not limited to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 16, or more). The cavities may be positioned in staggered rows,concentric circles, or any other arrangement.

The cavities may accept a sample, fluid or other substance directlytherein, or may accept a vessel and/or tip that may be configured toconfine or accept a sample, fluid, or other substance therein. Thecavities may be configured to accept a tip, such as a tip illustrated inFIG. 71, or any other tip and/or vessel described elsewhere herein. Thetest strip may optionally be a nucleic acid test strip, which may beconfigured to accept and support nucleic acid tips.

A cavity may have a tapered opening. In one example, a cavity mayinclude a top portion 7210 a, and a bottom portion 7210 b. The topportion may be tapered and may have an opening greater in diameter thanthe bottom portion.

In some embodiments, the cavity may be configured to accept a pipettenozzle for pick-up. One or more pipette nozzle may engage with one ormore cavity of the test strip. One, two, three, four, five, six or morepipette nozzles may simultaneously engage with corresponding cavities ofthe test strip. A tapered opening of the cavity may be useful for nozzlepick-up. The pipette nozzle may be press-fit into the cavity or mayinterface with the cavity in any other manner described herein.

One or more sample and/or reagent may be provided in a test strip. Thetest strips may have a narrow profile. A plurality of test strips may bepositioned adjacent to one another. In some instances, a plurality oftest strips adjacent to one another may form an array of cavities. Thetest strips may be swapped out for modular configurations. The teststrips and/or reagents may be movable independently of one another. Thetest strips may have different samples therein, which may need to bekept at different conditions and/or shuttled to different parts of thedevice on different schedules.

FIG. 73 shows another example of a test strip. The test strip may have abody 7300. The body may be formed from a single integral piece ormultiple pieces. The body may have a molded shape. The body may form aplurality of circular pieces 7310 a, 7310 b connected to one another, orvarious shapes connected to one another. The bodies of the circularpieces may directly connect to one another or one or more strip or spacemay be provided between the bodies.

The test strip may include one or more cavities 7330. In someembodiments, the cavities may be provided as a row in the body. Thecavities may optionally be provided in a straight row, in an array(e.g., m×n array where m, n are whole numbers greater than zeroincluding but not limited to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 16, or more). The cavities may be positioned in staggered rows,concentric circles, or any other arrangement.

The cavities may accept a sample, fluid or other substance directlytherein, or may accept a vessel and/or tip that may be configured toconfine or accept a sample, fluid, or other substance therein. Thecavities may be configured to accept a tip, such as a tip illustrated inFIG. 71, or any other tip and/or vessel described elsewhere herein. Thetest strip may optionally be a nucleic acid test strip, which may beconfigured to accept and support nucleic acid tips.

The test strip body 7330 may be molded around the cavities 7330. Forexample, if a cavity has a circular cross-section, the test strip bodyportion 7310 a, 7310 b around that cavity may have a circularcross-section. Alternatively, the test strip body need not match thecavity shape.

In some embodiments, the test strip may include an external pick-upreceptacle 7320. One or more pipette nozzle may engage with one or moreexternal pick-up receptacle of the test strip. One, two, three, four,five, six or more pipette nozzles may simultaneously engage withcorresponding pick-up receptacles of the test strip. A pick-upreceptacle may have one or more cavity 7340 or through-hole that may becapable of interfacing with a pipette nozzle. The pipette nozzle may bepress-fit into the cavity or may interface with the receptacle in anyother manner described herein.

One or more samples and/or reagents may be provided in a test strip. Theone or more sample may be directly within a cavity or may be provided intips and/or vessels that may be placed in a cavity of the test strip.The test strips may have a narrow profile. A plurality of test stripsmay be positioned adjacent to one another. In some instances, aplurality of test strips adjacent to one another may form an array ofcavities. The test strips may be swapped out for modular configurations.The test strips may be movable independently of one another. The teststrips and/or reagents may have different samples therein, which mayneed to be kept at different conditions and/or shuttled to differentparts of the device on different schedules.

Nucleic Acid Vessel/Tip

FIG. 24 shows an example of a vessel provided in accordance with anembodiment of the invention. In some instances, the vessel may be usedfor isothermal and non-isothermal nucleic acid assays (such as, withoutlimitation, LAMP, PCR, real-time PCR) or other nucleic acid assays.Alternatively, the vessel may be used for other purposes.

The vessel may include a body 2400 configured to accept and confine asample, wherein the body comprises an interior surface, an exteriorsurface, and open end 2410, and a closed end 2420. The vessel may beconfigured to engage with a pipette. The vessel may include a flexiblematerial 2430 extending through the cross-section of the vessel. Theflexible material may extend across the open end of the vessel.

The flexible material may or may not have a slit, hole, or other form ofopening. The flexible membrane may be configured to prevent fluid frompassing through the flexible membrane in the absence of an objectinserted through the slit. In some embodiments, the flexible materialmay be a membrane. The flexible material may be a septum formed of asilicon-based material, or any elastic or deformable material. In someembodiments, the flexible material may be a self-healing material. Anobject, such as a tip, may be inserted through the flexible material.The tip may be inserted through a slit or opening in the flexiblematerial or may penetrate the flexible material. FIG. 24 shows anexample of a tip inserted into a vessel, passing through the flexiblematerial, from an exterior view, and a cut-away view. The insertion ofthe tip may permit a sample to be dispensed to the vessel and/or beaspirated from the vessel through the tip. When the tip is removed, theflexible membrane may reseal or the slit may be sufficiently closed toprevent a fluid from passing through the flexible membrane.

The body of the vessel may have a first open end 2410 and a secondclosed end 2420. A cross-sectional dimension, such as a diameter, of thefirst end may be greater than the cross-sectional dimension of thesecond end. The closed end may have a tapered shape, rounded shape, or aflat shape.

In some embodiments, the body of the vessel may have a cylindricalportion 2440 of a first diameter having an open end 2442 and a closedend 2444, and a funnel shaped portion 2450 contacting the open end,wherein one end of the funnel shaped portion may contact the open endand may have the first diameter, and a second end 2452 of the funnelshaped portion may have a second diameter. In some embodiments, thesecond end of the funnel shaped portion may contact another cylindricalportion 2460 that has two open ends, and that may have the seconddiameter. In some embodiments, the second diameter may be greater thanthe first diameter. Alternatively, the first diameter may be greaterthan the second diameter. In some embodiments, the open end of thevessel body may be configured to engage with a removable cap 2470. Insome embodiments, an end of the additional cylindrical portion or asecond end of the funnel shaped portion may be configured to engage withthe cap.

In some embodiments, the vessel may also include a cap 2470. The cap maybe configured to contact the body at the open end of the body. In someembodiments, at least a portion of the cap may extend into the interiorof the body or may surround a portion of the body. Alternatively, aportion of the body may extend into the interior of the cap or maysurround a portion of the cap. The cap may have two or more ends. Insome embodiments, one, two or more of the ends may be open. For example,a cap may have a first end 2472 and a second end 2474. A passageway mayextend through the cap. The diameter of the cap may remain the samethroughout the length of the cap. Alternatively, the diameter of the capmay vary. For example, the end of the cap further from the body may havea smaller diameter than the end of the cap to be engaged with the body.

The flexible membrane 2430 may be provided within the body of thevessel. Alternatively, the flexible membrane may be provided within thecap of the vessel. The flexible membrane may be sandwiched between thebody and the cap of the vessel. In some instances, the flexible membranemay be provided both within the body and cap of the vessel, or multipleflexible membranes may be provided that may be distributed between thebody and cap of the vessel in any manner. In some embodiments, the bodymay comprise an interior portion through which the flexible materialextends, or the cap may comprise a passageway through which the flexiblematerial extends.

One or more tip may be inserted into the vessel. In some embodiments,the tip may be specially designed for insertion into a nucleic acidvessel. Alternatively, any of the tips described elsewhere herein may beinserted into the nucleic acid vessel. In some instances, a pipette tipmay be inserted into the nucleic acid vessel.

The tip 2480 may have a lower portion 2482 and an upper portion 2484.The lower portion may have an elongated shape. The lower portion mayhave a smaller diameter than the upper portion. One or more connectingfeature 2486 may be provided between the lower portion and the upperportion.

The lower portion of the tip may be inserted at least partially into thevessel. The tip may be inserted through the cap of the vessel and/orthrough the flexible material of the vessel. The tip may enter theinterior of the body of the vessel. The tip may pass through a slit oropening or of the flexible material. Alternatively, the tip may puncturethe flexible material.

In some embodiments, a tip and/or vessel may have any other type ofbarrier that may reduce contamination. The barrier may include aflexible material or membrane, film, oil (e.g., mineral oil), wax, gel,or any other material that may prevent a sample, fluid, or othersubstance contained within the tip and/or vessel from passing throughthe barrier. The barrier may prevent the substance within the tip and/orvessel from being contaminated by an environment, from aerosolizingand/or evaporating, and/or from contaminating other portions of thedevice. The barrier may permit a sample, fluid or other substance topass through the barrier only at desired conditions and/or times.

FIG. 25 shows an example of a vessel provided in accordance with anotherembodiment of the invention. In some instances, the vessel may be usedfor isothermal and non-isothermal nucleic acid assays (such as LAMP,PCR, real-time PCR) or other nucleic acid assays. Alternatively, thevessel may be used for other purposes. The vessel may or may not includefeatures or characteristics of the vessel described elsewhere herein.

The vessel may comprise a body 2500 configured to accept and confine asample, wherein the body comprises an interior surface, an exteriorsurface, a first end 2510, and a second end 2520. In some embodiments,one or more of the ends may be open. One or more of the ends may beclosed. In some embodiments, the first end may be open while the secondend may be closed. A passage may extend between the first and secondend.

The vessel may include a material 2530 extending across the passagecapable of having (1) a first state that is configured to prevent fluidfrom passing through the material in the absence of an object insertedinto the material, and a (2) second state that is configured to preventfluid and the object from passing through the material. The first statemay be a molten state and the second state may be a solid state. Forexample, when in the molten state, the material may permit a tip to passthrough, while preventing fluids from passing through. A fluid may bedispensed and/or aspirated through the tip passing through the material.The tip may be capable of being inserted through the material andremoved from the material while the material is in a molten state. Whenin the solid state, the material may be solid enough to prevent a tipfrom passing through and may prevent fluids from passing through.

In some embodiments, the material may be formed of wax. The material mayhave a selected melting point. For example, the material have a meltingpoint less than and/or equal to about 30 degrees C., 35 degrees C., 40degrees C., 45 degrees C., 50 degrees C., 55 degrees C., 60 degrees C.,65 degrees C., 70 degrees C., or 75 degrees C. The material may have amelting point between 50 and 60 degrees C. When the temperature of thematerial is sufficiently high, the material may enter a molten state.When the temperature of the material is brought sufficiently low, thematerial may solidify into a solid state.

When an object, such as a tip, is removed from the vessel through thematerial, a portion of the object may be coated with the material. Forexample, if a tip is inserted into molten wax, and then removed from thewax, the portion of the tip that was inserted into the wax may be coatedwith the wax when removed. This may advantageously seal the tip andreduce or prevent contamination. Also, the seal may prevent biohazardousor chemically hazardous material from escaping a vessel.

FIG. 25A shows an example of a nucleic acid amplification/wax assemblyvessel. The vessel may have a wax barrier 2530 and aqueous orlyophilized reagents 2550. The barrier may include molten wax that isplaced over reagents where it solidifies at shipping/storagetemperature.

FIG. 25B shows a second step where the vessel is heated to melt the waxand prepare for a sample. A pipette/nozzle 2540 may be used to place thevessel onto a heating block. Other mechanisms known in the art may beused to deliver heat to the wax. A wax barrier 2530 may be providedwhere the wax melts during the heating step. Aqueous or lyophilizedreagents 2550 may be provided beneath the wax barrier.

FIG. 25C shows the step of introducing a sample to the vessel. A tip2560, such as a pipette tip, may penetrate the molten wax barrier 2530.Aqueous or lyophilized reagents 2550 may be provided beneath thebarrier. The pipette tip may contain a DNA sample 2570 that may bedeposited beneath the wax layer. Depositing beneath the wax layer mayprevent contamination. The DNA containing sample may be deposited in thereagent layer. Optionally, when the tip is removed from the vessel, thetip may have a portion coated with wax.

FIG. 25D shows the step of amplification. The wax barrier 2530 may beprovided above the reagents and the sample layer 2550. The wax mayremain as a molten barrier during amplification. During the assay,amplification may take place under the wax layer. Turbidity or otherreadings may be taken during or after amplification to indicate thelevel of product.

FIG. 25E illustrates a step of post amplification wax solidification. Awax barrier 2530 may be provided above the reagent and sample layer2550. After assay readings are taken, the vessel may be cooled and thewax may resolidify, providing a containment barrier for the DNAgenerated by the nucleic acid amplification (e.g., PCR, real-time PCR,LAMP).

FIG. 25F shows the step of removal of the vessel. A pipette/nozzle 2540may be used to remove the fully contained used vessel. The vessel maycontain the wax barrier 2530 that has been solidified. The vessel mayalso contain the nucleic acid amplification product 2550, ready fordisposal. The pipette/nozzle may remove the vessel from a heat block ormay move the vessel to another portion of the device.

The pipette/nozzle may engage with the vessel through an open end of thevessel. In some embodiments, the pipette/nozzle may form a seal with thevessel. The pipette/nozzle may be press-fit to the vessel. Alternativelyadditional mechanisms may be used to allow the pipette/nozzle toselectively engage and/or disengage with the vessel.

Centrifugation Vessel/Tip

FIG. 26 shows an example of a vessel provided in accordance with anembodiment of the invention. In some instances, the vessel may be usedfor centrifugation. The vessel may be configured to be inserted into acentrifuge. Any centrifuge known in the art may be used. Examples ofcentrifuges are described in greater detail elsewhere herein. In oneembodiment, the vessel may be a centrifugation vessel. Alternatively,the vessel may be used for other purposes. In another non-limitingembodiment, the vessel has a tapered bottom in at least the interiorwall surfaces to allow the solids of the sample to aggregate. In thisexample, the length of the vessels is short enough so that the tips canbe inserted to the bottom of the vessel to resuspend the solutes asrequired. Optionally, the vessel volume is large enough to be able toprocess enough of the sample reducing sample processing times andreducing variability. Optionally, the vessel is narrow enough so thatvolumetric measurements of sample in the vessel are precise enough.

The vessel may comprise a body 2600 configured to accept and confine asample, wherein the body comprises an interior surface, an exteriorsurface, a first end 2608, and a second end 2610. In some embodiments,one or more of the ends may be open. One or more of the ends may beclosed. In some embodiments, the first end may be open while the secondend may be closed. A passage may extend between the first and secondend.

One or more end 2610 of a vessel may be round, tapered, flat, or haveany other geometry. In some embodiments, a cross-sectional dimension ofthe vessel, such as a diameter, may vary across the length of thevessel. In some instances, a lower portion 2620 of a vessel having aclosed end may have a smaller diameter than another upper portion 2630of the vessel closer to the open end. In some embodiments, one or moreadditional portion 2640 of the vessel may be provided which may belocated between the lower portion and the upper portion. In someembodiments, the diameter of the one or more additional portion may bebetween the sizes of the diameters of the lower portion and the upperportion. One or more funnel-shaped region 2650, step-shaped region, orridge 2660 may connect portions of different diameters. Alternatively,portions may transition gradually to have different diameters. In someembodiments, an open end of a vessel may have a greater cross-sectionaldimension than a closed end of a vessel.

Vessels interfacing with the centrifuge may be used for several purposesbeyond routine separation. Vessels interfacing with the centrifuge maybe designed for either separation or for specific assays. Examples ofassays that may be performed using the centrifuge include erythrocytesedimentation rate, red blood cell antibody screens, etc. Vessels usedfor these applications might be specialized with embeddedsensors/detectors, and ability to transmit data. Examples include tipswith in-built camera which can transmit images during red blood cellpacking. Centrifuge vessels may also be designed to be optimized forcentrifugal mixing, by using magnetic and/or non-magnetic beads.Centrifugation of cuvettes allows for forced flow inside small channels,which might be useful for applications such as fluid focusing andsize-based separations. Vessels may also be designed to process volumeswhich are much smaller than traditional centrifuges, where vessel designis critical to avoid destruction of fragile biological species such ascells. Centrifuge vessels may also be equipped with features to preventaerosolization without the need for capping the entire centrifuge.

In one embodiment, the vessel may be thought of as a two-piece part withthe top feature acting as a lid to prevent any fluid loss from thevessel in the form of aerosols. Alternatively, the vessel might beequipped with a septal duckbill valve to prevent aerosol leaks.

FIG. 26 also shows a tip provided in accordance with an embodiment ofthe invention. The tip may be used for dispensing and/or aspirating asample or other fluid from the vessel. The tip may be configured to beinserted at least partially into the vessel. In some embodiments, thetip may be a centrifuge extraction tip.

The tip may be configured to accept and confine a sample, wherein thetip comprises an interior surface, an exterior surface, a first end2666, and a second end 2668. In some embodiments, one or more of theends may be open. In some embodiments, the first and second ends may beopen. A passage may extend between the first and second end.

One or more end 2668 of a tip may be round, tapered, flat, or have anyother geometry. In some embodiments, a cross-sectional dimension of thetip, such as a diameter, may vary across the length of the tip. In someinstances, a lower portion 2670 of a tip at the second end may have asmaller diameter than another upper portion 2675 of the tip closer tothe first end. In some embodiments, one or more additional portion 2680of the tip may be provided which may be located between the lowerportion and the upper portion. In some embodiments, the diameter of theone or more additional portion may be between the sizes of the diametersof the lower portion and the upper portion. One or more funnel-shapedregion 2690, step-shaped region, or ridge 2695 may connect portions ofdifferent diameters. Alternatively, portions may transition gradually tohave different diameters. In some embodiments, a first end of a tip mayhave a greater cross-sectional dimension than a second end of a tip. Insome embodiments, the lower portion of the tip may be narrow and mayhave a substantially similar diameter throughout the length of the tip.

The tip may be configured to extend into the vessel through the open endof the vessel. The second end of the tip may be inserted into thevessel. The end of the tip having a smaller diameter may be insertedthrough an open end of the vessel. In some embodiments, the tip may beinserted fully into the vessel. Alternatively, the tip may be insertedonly partway into the vessel. The tip may have a greater height than thevessel. A portion of the tip may protrude outside of the vessel.

The vessel or the tip may comprise a protruding surface feature that mayprevent the second end of the tip from contacting the bottom of theinterior surface of the closed end of the vessel. In some embodiments,the protruding surface feature may be at or near the closed end of thevessel. In some embodiments, the protruding surface feature may belocated along the lower half of the vessel, lower ⅓ of the vessel, lower¼ of the vessel, lower ⅕ of the vessel, lower 1/10 of the vessel, lower1/20 of the vessel, or lower 1/50 of the vessel. The protruding surfacefeature may be located on an interior surface of the vessel.Alternatively, the protruding surface feature may be located on anexterior surface of the tip. In some instances, a protruding surfacefeature may be located on both the interior surface of the vessel andthe exterior surface of the tip.

In some embodiments, the protruding surface feature may include one ormore bump, ridge, or step. For example, a vessel may include the surfacefeatures integrally formed on the bottom interior surface of the vessel.The surface features may include one, two, three, four, five, six, ormore bumps on the bottom interior surface of the vessel. The surfacefeatures may be evenly spaced from one another. For example, the bumpsor other surface features may be provided in a radial pattern. The bumpsor other surface features may continuously or discontinuously encirclethe inner surface of the vessel, or the other surface of the tip.

Alternatively, the protruding surface features may be part of the shapeof the vessel or tip. For example, the vessel may be shaped with varyinginner diameters, and the tip may be shaped with varying outer diameters.In some embodiments, the inner surface of the vessel may form a step,upon which the tip may rest. The profile of the vessel and/or tip may beshaped so that based on the inner and outer cross-sectional dimensionsof the vessel and tip, the tip may be prevented from contacting thebottom of the vessel.

The vessel and/or tip may be shaped to prevent the tip from wigglingwithin the vessel when the tip has been inserted as far as it can go.Alternatively, the vessel and tip may be shaped to allow some wiggle. Insome embodiments, when the tip is inserted fully into the vessel, thetip may form a seal with the vessel. Alternatively, no seal need beformed between the tip and the vessel.

In some embodiments, the tip may be prevented from contacting the bottomof the vessel by a desired amount. This gap may enable fluid to freelyflow between the tip and the vessel. This gap may prevent choking offluid between the tip and the vessel. In some embodiments, the tip maybe prevented from contacting the bottom of the vessel to provide the tipat a desired height along the vessel. In some embodiments, one or morecomponents of a fluid or sample within the vessel may be separated andthe tip may be positioned to dispense and/or aspirate the desiredcomponents of the fluid or sample. For example, portions of the fluid orsample with a higher density may be provided toward the bottom of thevessel and portions with a lower density may be provided toward an upperportion of the vessel. Depending on whether the tip is to pick up ordeliver a fluid or sample to a higher density portion or lower densityportion, the tip may be located closer to the bottom and/or upperportion of the vessel respectively. In some embodiments, other featuresmay be provided to a centrifugation vessel and/or tip that may permitthe flow of fluid between the tip and the vessel at a desired heightalong the vessel. For example, the tip may comprise one or more opening,passageway, slit, channel, or conduit connecting the exterior surface ofthe tip to the passageway of the tip between the first and second ends.The opening may permit fluid flow, even if the end of the tip contactsthe bottom of the vessel. In some embodiments, a plurality of openingsmay be provided along the height of the tip. One or more opening may beprovided along the height of the tip to permit fluid flow at desiredheights within the vessel.

Tips may be configured to perform chromatography. In this process, themixture is dissolved in a fluid called the “mobile phase”, which carriesit through a structure holding another material called the “stationaryphase”. The various constituents of the mixture travel at differentspeeds, causing them to separate. The separation is based ondifferential partitioning between the mobile and stationary phases.Subtle differences in a compound's partition coefficient result indifferential retention on the stationary phase and thus changing theseparation. Tips may be configured to perform size exclusionchromatography, where molecules in solution are separated by their size,not by molecular weight. This can include gel filtration chromotography,gel permeation chromatography. Tips may be configured to enable themeasuring of mass-to-charge ratios of charged particles, therebyperforming mass spectrometry. Namely, the process ionizes chemicals togenerate charged molecules and then the ions are separated according totheir mass to charge ratio, possibly by an analyzer usingelectromagnetic fields. Tips may act as electrodes.

Systems and devices provided herein, such as point of service systems(including modules), are configured for use with vessels and tipsprovided in U.S. Patent Publication No. 2009/0088336 (“MODULARPOINT-OF-CARE DEVICES, SYSTEMS, AND USES THEREOF”), which is entirelyincorporated herein by reference.

Positive Displacement Tips

FIG. 27 also shows a tip 2700 provided in accordance with an embodimentof the invention. The tip may be used for dispensing and/or aspirating asample or other fluid from the vessel. The tip may be able to provideand/or pick up accurate and precise amounts of fluid, with highsensitivity. The tip may be configured to be inserted at least partiallyinto the vessel. In some embodiments, the tip may be a positivedisplacement tip such as but not limited to that shown in FIG. 14.

The tip may be configured to accept and confine a sample, wherein thetip comprises an interior surface, an exterior surface, a first end2702, and a second end 2704. In some embodiments, one or more of theends may be open. In some embodiments, the first and second ends may beopen. A passage may extend between the first and second end.

One or more end 2704 of a tip may be round, tapered, flat, or have anyother geometry. In some embodiments, a cross-sectional dimension of thetip, such as a diameter, may vary across the length of the tip. In someinstances, a lower portion 2710 of a tip at the second end may have asmaller diameter than another upper portion 2720 of the tip closer tothe first end. In some embodiments, one or more additional portion 2730of the tip may be provided which may be located between the lowerportion and the upper portion. In some embodiments, the diameter of theone or more additional portion may be between the sizes of the diametersof the lower portion and the upper portion. One or more funnel-shapedregion 2740, step-shaped region, or ridge 2750 may connect portions ofdifferent diameters. Alternatively, portions may transition gradually tohave different diameters. In some embodiments, a first end of a tip mayhave a greater cross-sectional dimension than a second end of a tip. Insome embodiments, the lower portion of the tip may be narrow and mayhave a substantially similar diameter throughout the length of the tip.

In some embodiments, a plunger 2760 may be provided that may be at leastpartially insertable within the positive displacement tip. In someembodiments, the tip may be dimensioned and/or shaped so that theplunger may be stopped from entering all the way to second end of thetip. In some embodiments, the tip may be stopped by an interior shelf2770. The tip may be preventing from entering a lower portion 2710 ofthe tip. An end 2765 of the plunger may be round, tapered, flat, or haveany other geometry.

The plunger may be configured to be movable within the tip. The plungermay move along the height of the tip. In some embodiments, the plungermay be movable to dispense and/or aspirate a desired volume of a sampleor other fluid.

The positive displacement tip may have an interior volume that may becapable of accepting any volume of fluid. For example, the positivedisplacement tip may have an interior volume that may contain less thanand/or equal to about 1 nL, 5 nL, 10 nL, 50 nL, 100 nL, 500 nL, 1 μL, 5μL, 8 μL, 10 μL, 15 μL, 20 μL, 30 μL, 40 μL, 50 μL, 60 μL, 70 μL, 80 μL,100 μL, 120 μL, 150 μL, 200 μL, 500 μL or any other volume describedelsewhere herein.

The tip may comprise one or more characteristics of the positivedisplacement tip as described elsewhere herein.

Additional Vessels/Tips

FIG. 28 shows an example of a well provided in accordance with anembodiment of the invention. The well may be an example of a vessel. Insome instances, the well may be used for various assays. The well may beconfigured to contain and/or confine one or more reagent. In someembodiments, one or more reaction may take place within the well.Alternatively, the well may be used for other purposes. In someembodiments, a plurality of wells may be provided. In some embodiments,384 wells may be provided. For example, the wells may be provided as oneor more rows, one or more columns, or an array. The wells may have 4.5μm diameters, and may be provided with 384 spacing. Alternatively, thewells may have any other spacing or size.

The well may comprise a body configured to accept and confine a sample,wherein the body comprises an interior surface, an exterior surface, afirst end 2806, and a second end 2808. In some embodiments, one or moreof the ends may be open. One or more of the ends may be closed. In someembodiments, the first end may be open while the second end may beclosed. A passage may extend between the first and second end.

One or more end 2808 of a well may be round, tapered, flat, or have anyother geometry. In some embodiments, a cross-sectional dimension of thevessel, such as a diameter, may vary across the length of the vessel.Alternatively, the cross-sectional dimension of the vessel need not varysubstantially. The vessel dimensions may transition gradually to havedifferent diameters. In some embodiments, an open end of a vessel mayhave a greater cross-sectional dimension than a closed end of a vessel.Alternatively, they open end and the closed end of the vessel may havesubstantially similar or the same cross-sectional dimension. In someembodiments, one or more end of the well may have a lip 2810, ridge, orsimilar surface feature. In some embodiments the lip may be provided ator near the open end of the well. The lip may be provided on an exteriorsurface of the well. In some embodiments, the lip may engage with ashelf that may support the well. In some embodiments, the lip may engagewith a cap that may cover the well. Capillaries and cuvettes are specialcases of fluid containment/processing units, since they are designed forspecific tasks. Capillaries in systems provided herein (e.g., bloodmetering capillaries) may utilize only capillary forces to transferfluid to specific locations. Cuvettes use a combination of capillaryand/or external forcing to transport fluids in specially designedchannels. Cuvettes and capillaries may be surface treated or finishedfor enhancing certain properties such as optical clarity, surfacetension, etc. or for addition of or coating with other substances suchas anti-coagulants, proteins, etc. Beads of different types may be usedin conjunction with specific vessels to further expand and/or enhanceprocessing in vessels. Examples include the following: a) Beads may beused to enhance mixing; b) Magnetic beads with coated antibody may beused. Bead separation is achieved by an external EM field; c)Non-magnetic beads may be used as an affinity column; d) Common beadssuch as polystyrene beads may be functionalized to capture specifictargets; and e) Long chain PEG beads may be used to make thread-likestructures.

FIG. 29 also shows a tip 2900 provided in accordance with an embodimentof the invention. The tip may be a bulk handling tip that may be usedfor dispensing and/or aspirating a sample or other fluid. The tip may beconfigured to be inserted at least partially into a vessel.Alternatively, the tip may be configured to dispense and/or aspirate asample or other fluid sample without being inserted into a vessel.

The tip may be configured to accept and confine a sample, wherein thetip comprises an interior surface, an exterior surface, a first end, anda second end. In some embodiments, one or more of the ends may be open.In some embodiments, the first and second ends may be open. A passagemay extend between the first and second end.

One or more end of a tip may be round, tapered, flat, or have any othergeometry. In some embodiments, a cross-sectional dimension of the tip,such as a diameter, may vary across the length of the tip. In someinstances, a lower portion 2910 of a tip at the second end may have asmaller diameter than another upper portion 2920 of the tip closer tothe first end. In some embodiments, one or more additional portion 2930of the tip may be provided which may be located between the lowerportion and the upper portion. In some embodiments, the diameter of theone or more additional portion may be between the sizes of the diametersof the lower portion and the upper portion. One or more funnel-shapedregion, step-shaped region, or ridge 2940 may connect portions ofdifferent diameters. Alternatively, portions may transition gradually tohave different diameters. In some embodiments, a first end of a tip mayhave a greater cross-sectional dimension than a second end of a tip. Insome embodiments, the lower portion of the tip may have a graduallychanging diameter. In some embodiments, a substantial difference indiameter may be provided along the length of the lower portion of thetip. A bulk handling tip may have a greater internal volume than one ormore of the other types of tips described herein.

FIG. 30 shows another example of a tip 3000 provided in accordance withan embodiment of the invention. The tip may be an assay tip configuredto provide a colorimetric readout (i.e., color tip) that may be used fordispensing and/or aspirating a sample or other fluid. The color tip maybe read using a detection system. The detection system may beincorporated from any of the embodiments described in greater detailelsewhere herein. The tip may be configured to be inserted at leastpartially into a vessel.

The tip may be configured to accept and confine a sample, wherein thetip comprises an interior surface, an exterior surface, a first end, anda second end. In some embodiments, one or more of the ends may be open.In some embodiments, the first and second ends may be open. A passagemay extend between the first and second end.

One or more end of a tip may be round, tapered, flat, or have any othergeometry. In some embodiments, a cross-sectional dimension of the tip,such as a diameter, may vary across the length of the tip. In someinstances, a lower portion 3010 of a tip at the second end may have asmaller diameter than another upper portion 3020 of the tip closer tothe first end. In some embodiments, one or more additional portion 3030of the tip may be provided which may be located between the lowerportion and the upper portion. In some embodiments, the diameter of theone or more additional portion may be between the sizes of the diametersof the lower portion and the upper portion. One or more funnel-shapedregion 3040, step-shaped region, or ridge 3050 may connect portions ofdifferent diameters.

Alternatively, portions may transition gradually to have differentdiameters. In some embodiments, a first end of a tip may have a greatercross-sectional dimension than a second end of a tip. In someembodiments, a relatively narrow lower portion of the tip may beprovided. The cross-sectional diameter of the lower portion need notchange or vary by a large amount. The lower portion of the tip may bereadable using a detection system. A detection system may be able todetect one or more signal pertaining to a sample or other fluid withinthe tip.

FIG. 31 provides a tip 3100 provided in accordance with anotherembodiment of the invention. The tip may be a blood tip that may be usedfor dispensing and/or aspirating a sample or other fluid. The tip may beconfigured to be inserted at least partially into a vessel. A tip may beconfigured as a “dip stick” that can be used to rapidly detect multipletargets, such as by using a thin pointed probe functionalized withreagents. In some embodiments, the fluid contained within the blood tipmay be blood.

The tip may be configured to accept and confine a sample, wherein thetip comprises an interior surface, an exterior surface, a first end, anda second end. In some embodiments, one or more of the ends may be open.In some embodiments, the first and second ends may be open. A passagemay extend between the first and second end.

One or more end of a tip may be round, tapered, flat, or have any othergeometry. In some embodiments, a cross-sectional dimension of the tip,such as a diameter, may vary across the length of the tip. In someinstances, a lower portion 3110 of a tip at the second end may have asmaller diameter than another upper portion 3120 of the tip closer tothe first end. In some embodiments, one or more additional portion 3130of the tip may be provided which may be located between the lowerportion and the upper portion. In some embodiments, the diameter of theone or more additional portion may be between the sizes of the diametersof the lower portion and the upper portion. One or more funnel-shapedregion 3140, step-shaped region, or ridge 3150 may connect portions ofdifferent diameters. Alternatively, portions may transition gradually tohave different diameters. In some embodiments, a first end of a tip mayhave a greater cross-sectional dimension than a second end of a tip. Insome embodiments, the lower portion of the tip may have a graduallychanging diameter. In some embodiments, a substantial difference indiameter may be provided along the length of the lower portion of thetip.

FIG. 32 provides a tip 3200 provided in accordance with anotherembodiment of the invention. The tip may be a current reaction tip thatmay be used for dispensing and/or aspirating a sample or other fluid.The tip may be configured to be inserted at least partially into avessel. In some embodiments, one or more reaction may take place withinthe tip.

The tip may be configured to accept and confine a sample, wherein thetip comprises an interior surface, an exterior surface, a first end, anda second end. In some embodiments, one or more of the ends may be open.In some embodiments, the tip may not fully enclose the passage. Forexample, an array of slotted pins can wick up fluids and deliver it tothe pipette by a blotting method. In some embodiments, the first andsecond ends may be open. A passage may extend between the first andsecond end.

One or more end of a tip may be round, tapered, flat, or have any othergeometry. In some embodiments, a cross-sectional dimension of the tip,such as a diameter, may vary across the length of the tip. In someinstances, a lower portion 3210 of a tip at the second end may have asmaller diameter than another upper portion 3220 of the tip closer tothe first end. In some embodiments, one or more additional portion 3230of the tip may be provided which may be located between the lowerportion and the upper portion. In some embodiments, the diameter of theone or more additional portion may be between the sizes of the diametersof the lower portion and the upper portion. One or more funnel-shapedregion, step-shaped region, or ridge 3240 may connect portions ofdifferent diameters. Alternatively, portions may transition gradually tohave different diameters. In some embodiments, a first end of a tip mayhave a greater cross-sectional dimension than a second end of a tip. Insome embodiments, the lower portion of the tip may have a graduallychanging diameter or may have substantially the same diameter.

Additional tips are provided in, for example, U.S. Patent PublicationNo. 2009/0088336 (“MODULAR POINT-OF-CARE DEVICES, SYSTEMS, AND USESTHEREOF”), which is entirely incorporated herein by reference.

Minitips

FIG. 33 shows an example of a minitip nozzle 3300 and a minitip 3310provided in accordance with an embodiment of the invention.

A minitip nozzle 3300 may be configured to interface with the minitip3310. In some embodiments, the minitip nozzle may connect to theminitip. The minitip may be attachable and detachable from the minitipnozzle. The minitip nozzle may be inserted at least partially into theminitip. The minitip nozzle may form a fluid-tight seal with theminitip. In some embodiments, the minitip nozzle may include a sealingo-ring 3320 or other sealing feature on its exterior surface. In otherembodiments, the minitip may include a sealing o-ring or other sealingfeature within its interior surface.

The minitip nozzle may be configured to interface with a fluid handlingdevice, such as a pipette. In some embodiments, the minitip nozzle maydirectly connect to a fluid handling device nozzle or orifice. Theminitip nozzle may form a fluid-tight seal with the fluid handlingdevice. In other embodiments, the minitip nozzle may connect to a tip orother intermediary structure that may be connected to the fluid handlingdevice.

FIG. 34 shows examples of minitips provided in accordance with anembodiment of the invention. For example, separate minitips may be usedto contain, dispense, and/or aspirate a volume less than and/or equal toabout 1 pL, 5 pL, 10 pL, 50 pL, 100 pL, 300 pL, 500 pL, 750 pL, 1 nL, 5nL, 10 nL, 50 nL, 75 nL, 100 nL, 125 nL, 150 nL, 200 nL, 250 nL, 300 nL,400 nL, 500 nL, 750 nL, 1 μL, 3 μL, 5 μL, 10 μL, or 15 μL in accordancewith an embodiment of the invention. The minitips may also be used forany other volume as described elsewhere herein.

A minitip may be configured to accept and confine a sample, wherein theminitip comprises an interior surface 3402, an exterior surface 3404, afirst end 3406, and a second end 3408. In some embodiments, one or moreof the ends may be open. In some embodiments, the first and second endsmay be open. A passage may extend between the first and second end.

One or more end 3408 of a minitip may be round, tapered, flat, or haveany other geometry. In some embodiments, a cross-sectional dimension ofthe minitip, such as a diameter, may vary across the length of the tip.In some instances, a lower portion 3410 of a tip at the second end mayhave a smaller diameter than another upper portion 3420 of the tipcloser to the first end. In some embodiments, one or more additionalportion of the tip may be provided which may be located between thelower portion and the upper portion. In some embodiments, the diameterof the one or more additional portion may be between the sizes of thediameters of the lower portion and the upper portion. Alternatively, nointermediate additional portion is provided between the lower and upperportions. One or more funnel-shaped region, step-shaped region, or ridge3430 may connect portions of different diameters. Alternatively,portions may transition gradually to have different diameters. In someembodiments, a first end of a tip may have a greater cross-sectionaldimension than a second end of a tip. In some embodiments, the lowerportion of the tip may have a gradually changing diameter or may havesubstantially the same diameter. The vessel may be covered by a rigid,and/or porous, and/or semi-permeable barrier in order to preventaerosolization, vaporization, etc. of the fluid, thereby preventing anycontamination of the device. Vessels may be designed with the ability toprocess small volumes (less than 10 uL) of fluid in POS devices, therebyreducing sample requirement. The vessel can be designed not only tocontain fluid, but also as to act as a location where unit operationsare carried out, including, but not limited to: separation, mixing,reactions, etc., involving small volumes of fluids. The vessel may bedesigned with special surface properties and/or features to enableexecution of special processes. De-centralizing unit operations inindividual vessels will result in reduced sample waste, lowerresource/lower consumption, and more efficient execution of chemistries.

Microcard

FIG. 35 provides an example of a microcard in accordance with anembodiment of the invention. The microcard may include one or moresubstrates 3500 configured to support one or more tips, which mayoptionally be microtips or vessels, herein used interchangeably. Thetips or vessels may have characteristics or the format of any other tipsor vessels described elsewhere herein. A microcard may be configured tosupport the performance or detection of multiple assays disclosedelsewhere herein in the card. Use of a microcard may, for example,permit the simultaneous performance or detection of multiple arrayedassays in small volumes or on a common support.

The microcard may optionally form a cartridge or be included within acartridge. The cartridge may be insertable and/or removable from asample processing device. The microcard may be insertable and/orremovable from the sample processing device.

The substrate may have a substantially planar configuration. In someembodiments, the substrate may have an upper surface and a lowersurface. The upper surface and lower surface may have a planarconfiguration. Alternatively, the upper and/or lower surface may have acurved surface, bent surface, surface with ridges or other surfacefeatures. The upper surface and opposing lower surface may be parallelto one another. Alternatively, upper and lower surfaces may have aconfiguration where they are not parallel to one another. In someembodiments, the planar substrate may have a plurality of depressions orcavities.

The substrate may have any shape known in the art. For example, thesubstrate may have a substantially square or rectangular shape.Alternatively, the substrate may have a circular, elliptical,triangular, trapezoidal, parallelogram, pentagonal, hexagonal,octagonal, or any other shape.

The substrate may have any lateral dimension (e.g., diameter, width,length). In some embodiments, one or more lateral dimension may be about0.1 mm, 0.5 mm, 1 mm, 5 mm, 7 mm, 1 cm, 1.5 cm, 2 cm, 2.5 cm, 3 cm, 3.5cm, 4 cm, 4.5 cm, 5 cm, 5.5 cm, 6 cm, 6.5 cm, 7 cm, 7.5 cm, 8 cm, 9 cm,10 cm, 11 cm, 12 cm, 13 cm, 15 cm, or 20 cm. The lateral dimensions maybe the same, or may vary.

The substrate may have any height (wherein height may be a dimension ina direction orthogonal to a lateral dimension). For example, the heightmay be less than or equal to about 0.1 mm, 0.5 mm, 1 mm, 1.5 mm, 2 mm,2.5 mm, 3 mm, 3.5 mm, 4 mm, 4.5 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 1 cm,1.2 cm, 1.5 cm, 2 cm, 2.5 cm, 3 cm, 4 cm, or 5 cm.

The substrate may be formed from any material. The substrate may beformed of a rigid, semi-rigid or flexible material. In some embodiments,the substrate include a metal, such as aluminum, steel, copper, brass,gold, silver, iron, titanium, nickel, or any alloy or combinationthereof, or any other metal described elsewhere herein. In otherembodiments, the substrate may include silicon, plastic, rubber, wood,graphite, diamond, resin, or any other material, including but notlimited to those described elsewhere herein. One or more surface of thesubstrate may or may not be coated with a material. For example, one ormore portion of the cavity may be coated with a rubbery material thatmay grip the vessels and/or tips and prevent them from slipping out.

The substrate may be substantially solid or hollow. The substrate may beformed from a solid material with one or more cavities provided therein.Alternatively, the substrate may have a shell-like structure. Thesubstrate may include a cage-like or mesh-like structure. The substratemay include one or more components that may link cavities together.Linking components may include bars, chains, springs, sheets, blocks, orany other components.

The substrate may be configured to support one or more tips or vessels.The substrate 3500 may contain one or more cavity 3510 configured toaccept one or more tips or vessels. The cavities may have anyarrangement on the substrate. For example, the cavities may form one ormore rows and/or one or more columns. In some embodiments, the cavitiesmay form an m×n array where m, n are whole numbers. Alternatively, thecavities may form staggered rows and/or columns. The cavities may formstraight lines, curved lines, bent lines, concentric patterns, randompatterns, or have any other configuration known in the art.

Any number of cavities may be provided on a substrate. For example,greater than and/or equal to about 1 cavity, 4 cavities, 6 cavities, 10cavities, 12 cavities, 24 cavities, 25 cavities, 48 cavities, 50cavities, 75 cavities, 96 cavities, 100 cavities, 125 cavities, 150cavities, 200 cavities, 250 cavities, 300 cavities, 384 cavities, 400cavities, 500 cavities, 750 cavities, 1000 cavities, 1500 cavities, 1536cavities, 2000 cavities, 3000 cavities, 3456 cavities, 5000 cavities,9600 cavities, 10000 cavities, 20000 cavities, 30000 cavities, or 50000cavities may be provided on a single substrate of the microcard.

The cavities may all have the same dimensions and/or shapes or may vary.In some embodiments, a cavity may extend partway into the substratewithout breaking through the substrate. A cavity may have an interiorwall and a bottom surface. Alternatively, the cavity may extend throughthe substrate. The cavity may or may not have a bottom surface orpartial bottom surface or shelf.

The cavities may have any geometry. For example, a cross-sectional shapeof a cavity may include circles, ellipses, triangles, quadrilaterals(e.g., squares, rectangles, trapezoids, parallelograms), pentagons,hexagons, octagons or any other shape. The cross-sectional shape of thecavity may remain or the same or vary along the height of the cavity.The cross-sectional shape of the cavity may be the same for all cavitieson a substrate, or may vary from cavity to cavity on the substrate. Thecross-sectional shapes of the cavity may or may not be complementary tothe exterior shape of a vessel and/or tip. The cavities may be formed aswells, or may be formed from cuvettes, or may have formats similar tomicrotiter plates.

The cavity may have any cross-sectional dimension (e.g., diameter,width, or length). For example, the cross-sectional dimension may begreater than or equal to about 0.1 mm, 0.5 mm, 1 mm, 1.5 mm, 2 mm, 2.5mm, 3 mm, 3.5 mm, 4 mm, 4.5 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 1 cm, 1.2cm, 1.5 cm, 2 cm, or 3 cm. The cross-sectional dimension may refer to aninner dimension of the cavity. The cross-sectional dimension may remainthe same throughout the height of the cavity or may vary. For example,an open upper portion of the cavity may have a greater cross-sectionaldimension than a closed bottom.

The cavity may have any height (wherein height may be a dimension in adirection orthogonal to a cross-sectional dimension). For example, theheight may be less than or equal to about 0.1 mm, 0.5 mm, 1 mm, 1.5 mm,2 mm, 2.5 mm, 3 mm, 3.5 mm, 4 mm, 4.5 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm,1 cm, 1.2 cm, 1.5 cm, 2 cm, 3 cm, 4 cm, or 5 cm. The height of thecavity may be less than the thickness of the substrate. Alternatively,the height of the cavity may be equal to the thickness of the substratewhen the cavity extends all the way through.

The bottoms of the cavities may have any shape. For example, the bottomsof the cavities may be rounded, flat, or tapered. The bottoms of thecavities may be complementary to a portion of one or more vessels and/ortips. The bottoms of the cavities may be complementary to a lowerportion of one or more vessels and/or tips. In some embodiments, thecavities may contain one or more surface feature that may permit thecavities to engage with a plurality of vessels and/or microtips.Different vessels and/or tips may engage different surfaces or portionsof the cavities. Alternatively, the cavities may be shaped to acceptparticular vessels and/or tips.

The interior of the cavity may have a volume of about 1,000 μL or less,500 μL or less, 250 μL or less, 200 μL or less, 175 μL or less, 150 μLor less, 100 μL or less, 80 μL or less, 70 μL or less, 60 μL or less, 50μL or less, 30 μL or less, 20 μL or less, 15 μL or less, 10 μL or less,8 μL or less, 5 μL or less, 1 μL or less, 500 nL or less, 300 nL orless, 100 nL or less, 50 nL or less, 10 nL or less, or 1 nL or less.

The cavities may be shaped to receive particular tips or vessels. Insome embodiments, the cavities may be shaped to receive a plurality ofdifferent types of tips and/or vessels. The cavity may have an internalsurface. At least a portion of the internal surface may contact a vesseland/or tip. In one example, the cavity may have one or more shelf orinternal surface features that may permit a first vessel/tip having afirst configuration to fit within the cavity and a second vessel/tiphaving a second configuration to fit within the cavity. The first andsecond vessels/tips having different configurations may contactdifferent portions of the internal surface of the cavity. In someembodiments, cavities of a microcard are configured to interface withminuatured tips (e.g. which can support a volume of no greater than, forexample, 20, 10, 5, 3, 2, 1, 0.5, or 0.1 microliter).

In some embodiments, the cavities may accept one or more vessels and/ormicrotips. The vessels and/or tips may be snap fitted into the cavities.Alternatively, the vessels and/or microtips may slide in and out of thecavity smoothly, may be press-fit into the cavities, may be twisted intothe cavity, or may have any other interaction with the cavities.

Alternatively, the cavities need not accept vessel and/or tips. Thecavities themselves may form vessels that may contain and/or confine oneor more fluid. For example, the cavities themselves may be a samplecontainer or may contain any other fluid, including reagents. Thecavities may be designed so that light does not pass through thecavities. In some instances, fluids or selected chemicals do not passthrough the cavity walls.

The cavities may all have openings on the same side of the substrate. Insome embodiments, the cavities may all open up to an upper surface ofthe substrate. Alternatively, some cavities may open to a lower surfaceof the substrate and/or a side surface of the substrate.

In some embodiments, the cavities may be formed using lithographictechniques, etching, laser etching, drilling, machining, or any othertechnique known in the art. The cavities may be cut into the substrate.

One or more vessels and/or microtips may be inserted into the cavities.An individual cavity may be configured to accept a single vessel and/ortip. Alternatively, an individual cavity may be configured to accept aplurality of vessels and/or microtips simultaneously. The cavities mayall be filled with vessels and/or microtips, or some cavities may bevacant.

Vessels and/or tips may be at least partially inserted into thecavities. The vessels and/or tips may extend beyond a surface of thesubstrate. For example, if the cavities of the substrate have an openingon an upper surface of the substrate, the vessels and/or tips may extendbeyond the upper surface of the substrate. At least a portion of avessel and/or microtip may protrude from the substrate. Alternatively, aportion of a vessel and/or tip does not protrude from the substrate. Thedegree to which a vessel and/or tip protrudes from the substrate maydepend on the type of vessel and/or tip, or cavity configuration.

In some alternate embodiments, a vessel and/or microtip may extend allthe way through a substrate. A vessel and/or microtip may extend abovetwo or more surfaces of the substrate. In some embodiments, a vesseland/or tip may extend at least partially beyond a lower surface of thesubstrate.

The vessels and/or microtips may be supported by the substrate so thatthey are parallel to one another. For example, the vessels and/or tipsmay all have a vertical alignment. The vessels and/or microtips may bealigned to be orthogonal to a planar surface of the substrate. Thevessel and/or tips may be orthogonal to a top surface and/or bottomsurface of the substrate. Alternatively the vessel and/or tips need notbe parallel to one another.

In some embodiments, each cavity may have a vessel and/or tip providedtherein. Alternatively, some cavities may be intentionally left open.One or more controller may track whether a cavity is occupied or empty.One or more sensor may determine if a cavity is occupied or empty.

The vessels and/or tips may be selectively placed and/or removed fromthe substrate. A vessel and/or microtip may be removed from a cavity ofa substrate to another portion of the device, or to another cavity ofthe substrate. A vessel and/or microtip may be placed in a cavity of thesubstrate from another portion of the device, or from another cavity ofthe substrate. Positions of vessels and/or microtips on a substrate maybe modified or exchanged. In some embodiments, each of the cavities maybe individually addressable. Each of the vessels and/or tips may beindividually addressable and/or movable. The vessels and/or microtipsmay be addressed and/or moved independently of one another. For example,a single vessel and/or microtip may be addressed and/or moved relativeto the other vessels and/or microtips. A plurality of vessels and/ormicrotips may be moved simultaneously. In some instances, a singlevessel and/or microtip may be moved at a time. The individual vesselsand/or microtips may be movable relative to one another and/or thecavities.

A vessel and/or tip may be removed and/or placed from a substrate usinga fluid handling device. A vessel and/or tip may be removed and/orplaced using another automated process not requiring human interaction.Alternatively, a vessel and/or tip can be manually removed and/orplaced. The vessel and/or tip may be individually moved in an automatedor manual process.

A microcard may include a plurality of vessels and/or tips of differenttypes. A microcard may include at least two, at least three, at leastfour, at least five, or at least six or more different types of vesselsand/or tips. Alternatively, a microcard may include all of the sametypes of vessels and/or tips. The microcard may include one or morevessels and/or tips selected from the following: nucleic acid vessel,nucleic acid tip, centrifugation vessel, centrifugation tip, positivedisplacement tip, well, bulk handling tip, color tip, blood tip, currentreaction tip, 3 μL minitip, 5 μL minitip, 10 μL minitip, or 15 μLminitip, or any other tips/vessels or combinations thereof. Themicrocard may include one or more vessels and/or tips configured toperform one or more of the following assays: immunoassay, nucleic acidassay, receptor-based assay, cytometric assay, colorimetric assay,enzymatic assay, electrophoretic assay, electrochemical assay,spectroscopic assay, chromatographic assay, microscopic assay,topographic assay, calorimetric assay, turbidimetric assay,agglutination assay, radioisotope assay, viscometric assay, coagulationassay, clotting time assay, protein synthesis assay, histological assay,culture assay, osmolarity assay, and/or other types of assays orcombinations thereof. One, two, three, four, five, six, or more of theassays may be supported by the vessels and/or tips supported by thesubstrate.

In some embodiments, microcards are configured for the performance ofimmunoassays. A microcard may contain different antibody-labeled beadsin different cavities of the microcard. In some embodiments, cavitiescontaining antibody-labeled beads do not contain vessels or tips. Thebeads of the antibody-labeled beads may of any type, including magneticbeads. While remaining in the cavities of the microcard, theantibody-labeled beads may be incubated with sample, washed, mixed withdetection reagents, and brought into proximity with a detection unit, inorder to detect whether the relevant analyte was in the sample.

Assay Units

In accordance with an embodiment of the invention, an assay station, orany other portion of a module or device, may include one or more assayunits. An assay unit may be configured to perform a biological orchemical reaction that yields a detectable signal indicative of thepresence or absence of one or more analyte, and/or a concentration of aone or more analyte. An assay unit may be configured to run an assay,which may include any type of assay as described elsewhere herein. Theassay may occur within the assay unit.

A detectable signal may include an optical signal, visible signal,electrical signal, magnetic signal, infrared signal, thermal signal,motion, weight, or sound.

In some embodiments, a plurality of assay units may be provided. In someembodiments, one or more row of assay units, and/or one or more columnof assay units may be provided. In some embodiments, an m×n array ofassay units may be provided, wherein m, n are whole numbers. The assayunits may be provided in staggered rows or columns from each other. Insome embodiments, they may have any other configuration.

Any number of assay units may be provided. For example there may be morethan and/or equal to about 1, 2, 3, 4, 5, 8, 10, 15, 20, 25, 30, 40, 50,60, 70, 80, 90, 100, 120, 150, 175, 200, 250, 300, 400, 500, or 1000assay units.

Assay units may be provided in a cartridge, card, or have any othersupporting structure. The assay units may have the same orientation.Alternatively, assay units may have different orientations. In someexamples, assay units may be kept at a vertical orientation. In otherexamples, assay units may have horizontal or vertical orientations, orany other angle of orientation. The assay units may remain the same ormay vary over time.

The assay units may be fluidically isolated or hydraulically independentfrom one another. The assay units may contain and/or confine samples orother fluids that may be in fluid isolation from one another. Thesamples and/or other fluids contained within the assay units may be thesame, or may vary from unit to unit. The system may be capable oftracking what each assay unit contains. The system may be capable oftracking the location and history of each assay unit.

The assay units may be independently movable relative to one another, oranother portion of the device or module. Thus, the fluids and/or samplescontained therein may be independently movable relative to one anotheror other portions of the device or module. An assay unit may beindividually addressable. The location of each assay unit may betracked. An assay unit may be individually selected to receive and/orprovide a fluid. An assay unit may be individually selected to transporta fluid. Fluid may be individually provided to or removed from an assayunit. Fluid may be individually dispensed and/or aspirated using theassay unit. An assay unit may be independently detectable.

Any description herein of individual assay units may also apply togroups of assay units. A group of assay units may include one, two, ormore assay units. In some embodiments, assay units within a group may bemoved simultaneously. The location of groups of assay units may betracked. Fluids may be simultaneously delivered and/or aspirated fromone or more group of assay units. Detection may occur simultaneously toassay units within one or more groups of assay units.

The assay units may have the form or characteristics of any of the tipsor vessels as described elsewhere herein. For example, an assay unit canbe any of the tips or vessels described herein. Any description hereinof assay units may also apply to tips or vessels, or any description oftips or vessels may also apply to the assay units.

In some embodiments, an assay unit may be an assay tip. An assay tip mayhave a first end and a second end. The first end and second end may beopposing one another. The first end and/or the second end may be open orclosed. In some embodiments, both the first and second ends may be open.In alternate embodiments, the assay unit may have three, four, or moreends.

The assay tip may have an interior surface and an exterior surface. Apassageway may connect the first and second ends of the assay tip. Thepassageway may be a conduit or channel. The first and second ends of theassay tip may be in fluid communication with one another. The diameterof the first end of the assay tip may be greater than the diameter ofthe second end of the assay tip. In some embodiments, the outer diameterof the first end of the assay tip may be greater than the outer diameterof the second end of the assay tip. An inner diameter of the first endof the assay tip may be greater than the inner diameter of the secondend of the assay tip. Alternatively, a diameter of the assay tip may bethe same at the first and second ends. In some embodiments, the secondend may be held below the first end of the assay tip. Alternatively therelative positions of the first and second ends may vary.

As previously described regarding tips and/or vessels, an assay unit maybe picked up using a fluid handling device. For example, a pipette orother fluid handling device may connect to the assay unit. A pipettenozzle or orifice may interface with an end of the assay unit. In someembodiments, a fluid-tight seal may be formed between the fluid handlingdevice and the assay unit. An assay unit may be attached to and/ordetached from the fluid handling device. Any other automated device orprocess may be used to move or manipulate an assay unit. An assay unitmay be moved or manipulated without the intervention of a human.

A fluid handling device or any other automated device may be able topick up or drop off an individual assay unit. A fluid handling device orother automated device may be able to simultaneously pick up or drop offa plurality of assay units. A fluid handling device or other automateddevice may be able to selectively pick up or drop off a plurality ofassay units. In some embodiments, a fluid handling device may be able toselectively aspirate and/or dispense a sample using one, two or moreassay units. Any description of fluid handling systems as describedpreviously herein may apply to the assay units.

In one embodiment, an assay unit may be formed from molded plastic. Theassay unit may be either commercially available or can be made by custommanufacturing with precise shapes and sizes. The units can be coatedwith capture reagents using method similar to those used to coatmicrotiter plates but with the advantage that they can be processed inbulk by placing them in a large vessel, adding coating reagents andprocessing using sieves, holders, and the like to recover the pieces andwash them as needed. In some embodiments, the capture reagents may beprovided on an interior surface of the assay units.

An assay unit can offer a rigid support on which a reactant can beimmobilized. The assay unit is also chosen to provide appropriatecharacteristics with respect to interactions with light. For example,the assay unit can be made of a material, such as functionalized glass,Si, Ge, GaAs, GaP, SiO₂, SiN₄, modified silicon, or any one of a widevariety of gels or polymers such as (poly)tetrafluoroethylene,(poly)vinylidenedifluoride, polystyrene, polycarbonate, polypropylene,PMMA, ABS, or combinations thereof. In an embodiment, an assay unit maycomprise polystyrene. Other appropriate materials may be used inaccordance with the present invention. Any of the materials describedhere, such as those applying to tips and/or vessels may be used to forman assay unit. A transparent reaction site may be advantageous. Inaddition, in the case where there is an optically transmissive windowpermitting light to reach an optical detector, the surface may beadvantageously opaque and/or preferentially light scattering.

A reactant may be immobilized at the capture surface of an assay unit.In some embodiments, the capture surface is provided on an interiorsurface of the assay unit. In one example, the capture surface may beprovided in a lower portion of an assay tip. The reagent can be anythinguseful for detecting an analyte of interest in a sample of bodily fluid.For instance, such reactants include, without limitation, nucleic acidprobes, antibodies, cell membrane receptors, monoclonal antibodies andantisera reactive with a specific analyte. Various commerciallyavailable reactants such as a host of polyclonal and monoclonalantibodies specifically developed for specific analytes can be used.

One skilled in the art will appreciate that there are many ways ofimmobilizing various reactants onto a support where reaction can takeplace. The immobilization may be covalent or noncovalent, via a linkermoiety, or tethering them to an immobilized moiety. Non-limitingexemplary binding moieties for attaching either nucleic acids orproteinaceous molecules such as antibodies to a solid support includestreptavidin or avidin biotin linkages, carbamate linkages, esterlinkages, amide, thiolester, (N)-functionalized thiourea, functionalizedmaleimide, amino, disulfide, amide, hydrazone linkages, and amongothers. In addition, a silyl moiety can be attached to a nucleic aciddirectly to a substrate such as glass using methods known in the art.Surface immobilization can also be achieved via a Poly-L Lysine tether,which provides a charge-charge coupling to the surface.

The assay units can be dried following the last step of incorporating acapture surface. For example, drying can be performed by passiveexposure to a dry atmosphere or via the use of a vacuum manifold and/orapplication of clean dry air through a manifold.

In some embodiments, rather than using a capture surface on the assayunit, beads or other substrates may be provided to the assay units withcapture surfaces provided thereon. One or more free-flowing substratemay be provided with a capture surface. In some embodiments, thefree-flowing substrate with a capture surface may be provided within afluid. In some embodiments, a bead may be magnetic. The bead may becoated with one or more reagents as known in the art. A magnetic beadmay be held at a desired location within the assay unit. The magneticbead may be positioned using one or more magnet.

Beads may be useful for conducting one or more assay, including but notlimited to immunoassay, nucleic acid assay, or any of the other assaysdescribed elsewhere herein. The beads may be used during a reaction(e.g., chemical, physical, biological reaction). The beads may be usedduring one or more sample preparation step. The beads may be coated withone or more reagent. The beads themselves may be formed of reagents. Thebeads may be used for purification, mixing, filtering, or any otherprocesses. The beads may be formed of a transparent material,translucent material, and/or opaque material. The beads may be formed ofa thermally conductive or thermally insulative material. The beads maybe formed of an electrically conductive or electrically insulativematerial. The beads may accelerate a sample preparation and/or assaystep. The beads may provide an increased surface area that may reactwith one or more sample or fluid.

In alternate embodiments, beads or other solid materials may be providedto the assay units. The beads may be configured to dissolve undercertain conditions. For example, the beads may dissolve when in contactwith a fluid, or when in contact with an analyte or other reagents. Thebeads may dissolve at particular temperatures.

The beads may have any size or shape. The beads may be spherical. Thebeads may have a diameter of less than or equal to about 1 nm, 5 nm, 10nm, 50 nm, 100 nm, 200 nm, 300 nm, 500 nm, 750 nm, 1 μm, 2 μm, 3 μm, 5μm, 10 μm, 20 μm, 50 μm, 100 μm, 200 μm, 300 μm, 400 μm, 500 μm, 600 μm,700 μm, 800 μm, 900 μm, 1 mm, 1.2 mm, 1.5 mm, 2 mm, 2.5 mm, 3 mm, 4 mm,or 5 mm. The beads may be of the same size or differing sizes. The beadsmay include microparticles or nanoparticles.

Any description of beads in the assay unit, processing unit, and/orreagent unit may be applied to beads located anywhere in the device.Beads may be stored and/or used in any tips/vessels (including thosedescribed herein), cuvettes, capillaries, channels, tanks, reservoirs,chambers, conduits, tubes, pipes, on surfaces, or any other location.Beads may be provided in a fluid, or may be separate from a fluid.

A reaction site may be provided within an assay unit. In someembodiments, a reaction site may be provided on a surface, such as theinterior surface, of the assay unit. The reaction site may be providedwithin a fluid contained by the assay unit. The reaction site may be ona substrate within the assay unit. The reaction site may be on thesurface of a substrate free-floating within the assay unit. The reactionsite may be a substrate within the assay unit.

An assay unit may have any dimension, including those describedelsewhere herein for tips and/or vessels. The assay unit may be capableof containing and/or confining a small volume of sample and/or otherfluid, including volumes mentioned elsewhere herein.

An assay unit may be picked up and/or removed from a fluid handlingmechanism. For example, an assay tip or other assay unit may be pickedup by a pipette nozzle. The assay tip or other assay unit may be droppedoff by a pipette nozzle. In some embodiments, assay units may beselectively individually picked up and/or dropped off. One or more groupof assay units may be selectively picked up and/or dropped off. An assayunit may be picked up and/or dropped off using an automated mechanism.An assay unit may be picked up and/or dropped off without requiringhuman intervention. A pipette may pick up and/or drop off an assay unitin accordance with descriptions provided elsewhere herein.

An assay unit may be moved within a device and/or module using a fluidhandling mechanism. For example, an assay tip or other assay unit may betransported using a pipette head. The assay tip or other assay unit maybe transported in a horizontal direction and/or vertical direction. Theassay tip and/or assay unit may be transported in any direction. Theassay unit may be moved individually using the fluid handling mechanism.One or more groups of assay units may be simultaneously moved using thefluid handling mechanism.

An assay unit may be shaped and/or sized to permit detection by adetection unit. The detection unit may be provided external to, inside,or integrated with the assay unit. In one example, the assay unit may betransparent. The assay unit may permit the detection of an opticalsignal, audio signal, visible signal, electrical signal, magneticsignal, motion, acceleration, weight, or any other signal by a detectionunit.

A detector may be capable of detecting signals from individual assayunits. The detector may differentiate signals received from each of theindividual assay units. The detector may individually track and/orfollow signals from each of the individual assay units. A detector maybe capable of simultaneously detecting signals from one or more groupsof assay units. The detector may track and/or follow signals from theone or more groups of assay units.

An assay unit may be formed from any material. An assay unit may beformed from any material including those described for tips and/orvessels elsewhere herein. An assay unit may be formed from a transparentmaterial.

Processing Units

In accordance with an embodiment of the invention, a preparation stationand/or assay station, or any other portion of a module or device, mayinclude one or more processing units. A processing unit may beconfigured to prepare a sample for the performance and/or to perform abiological or chemical reaction that yields a detectable signalindicative of the presence or absence of one or more analyte, and/or aconcentration of a one or more analyte. The processing unit may be usedfor preparing an assay sample or performing any other process withrespect to the sample or related reagents, as provided in one or moresample preparation or processing steps as described elsewhere herein.The processing unit may have one or more characteristics of an assayunit as described elsewhere herein. A processing unit may function as anassay unit as described elsewhere herein.

A detectable signal may include an optical signal, visible signal,electrical signal, magnetic signal, infrared signal, thermal signal,motion, weight, or sound.

In some embodiments, a plurality of processing units may be provided. Insome embodiments, one or more row of processing units, and/or one ormore column of processing units may be provided. In some embodiments, anm×n array of processing units may be provided, wherein m, n are wholenumbers. The processing units may be provided in staggered rows orcolumns from each other. In some embodiments, they may have any otherconfiguration.

Any number of processing units may be provided. For example there may bemore than and/or equal to about 1, 2, 3, 4, 5, 8, 10, 15, 20, 25, 30,40, 50, 60, 70, 80, 90, 100, 120, 150, 175, 200, 250, 300, 400, 500, or1000 processing units.

Processing units may be provided in a cartridge, card, or have any othersupporting structure. The processing units may have the sameorientation. Alternatively, processing units may have differentorientations. In some examples, processing units may be kept at avertical orientation. In other examples, processing units may havehorizontal or vertical orientations, or any other angle of orientation.The processing units may remain the same or may vary over time.

In some cases, a pipette, tip, or both may be integrated with acartridge or card. In some cases, tips or pipettes, or components oftips or pipettes, are integrated with cartridges or cards.

The processing units may be fluidically isolated or hydraulicallyindependent from one another. The processing units may contain and/orconfine samples or other fluids that may be in fluid isolation from oneanother. The samples and/or other fluids contained within the processingunits may be the same, or may vary from unit to unit. The system may becapable of tracking what each processing unit contains. The system maybe capable of tracking the location and history of each processing unit.

The processing units may be independently movable relative to oneanother, or another portion of the device or module. Thus, the fluidsand/or samples contained therein may be independently movable relativeto one another or other portions of the device or module. A processingunit may be individually addressable. The location of each processingunit may be tracked. A processing unit may be individually selected toreceive and/or provide a fluid. A processing unit may be individuallyselected to transport a fluid. Fluid may be individually provided to orremoved from a processing unit. Fluid may be individually dispensedand/or aspirated using the processing unit. A processing unit may beindependently detectable.

Any description herein of individual processing units may also apply togroups of processing units. A group of processing units may include one,two, or more processing units. In some embodiments, processing unitswithin a group may be moved simultaneously. The location of groups ofprocessing units may be tracked. Fluids may be simultaneously deliveredand/or aspirated from one or more group of processing units. Detectionmay occur simultaneously to processing units within one or more groupsof processing units.

The processing units may have the form or characteristics of any of thetips or vessels as described elsewhere herein. For example, a processingunit can be any of the tips or vessels described herein. Any descriptionherein of processing units may also apply to tips or vessels, or anydescription of tips or vessels may also apply to the processing units.

In some embodiments, a processing unit may be a processing tip. Aprocessing tip may have a first end and a second end. The first end andsecond end may be opposing one another. The first end and/or the secondend may be open or closed. In some embodiments, both the first andsecond ends may be open. In alternate embodiments, the processing unitmay have three, four, or more ends.

The processing tip may have an interior surface and an exterior surface.A passageway may connect the first and second ends of the processingtip. The passageway may be a conduit or channel. The first and secondends of the processing tip may be in fluid communication with oneanother. The diameter of the first end of the processing tip may begreater than the diameter of the second end of the processing tip. Insome embodiments, the outer diameter of the first end of the processingtip may be greater than the outer diameter of the second end of theprocessing tip. An inner diameter of the first end of the processing tipmay be greater than the inner diameter of the second end of theprocessing tip. Alternatively, a diameter of the processing tip may bethe same at the first and second ends. In some embodiments, the secondend may be held below the first end of the processing tip. Alternativelythe relative positions of the first and second ends may vary.

In some embodiments, a processing unit may be a vessel. A processingunit may have a first end and a second end. The first end and second endmay be opposing one another. The first end and/or the second end may beopen or closed. In some embodiments, the second end may be held belowthe first end of the processing unit. Alternatively the relativepositions of the first and second ends may vary. An open end of theprocessing unit may be oriented upwards, or may be held higher than aclosed end.

In some embodiments, a processing unit may have a cap or closure. Thecap or closure may be capable of blocking an open end of the processingunit. The cap or closure may be selectively applied to close or open theopen end of the processing unit. The cap or closure may have one or moreconfiguration as illustrated elsewhere herein or as known in the art.The cap or closure may form an airtight seal that may separate thecontents of the reagent unit from the ambient environment. The cap orclosure may include a film, oil (e.g., mineral oil), wax, or gel.

As previously described regarding tips and/or vessels, a processing unitmay be picked up using a fluid handling device. For example, a pipetteor other fluid handling device may connect to the processing unit. Apipette nozzle or orifice may interface with an end of the processingunit. In some embodiments, a fluid-tight seal may be formed between thefluid handling device and the processing unit. A processing unit may beattached to and/or detached from the fluid handling device. Any otherautomated device or process may be used to move or manipulate aprocessing unit. A processing unit may be moved or manipulated withoutthe intervention of a human.

A fluid handling device or any other automated device may be able topick up or drop off an individual processing unit. A fluid handlingdevice or other automated device may be able to simultaneously pick upor drop off a plurality of processing units. A fluid handling device orother automated device may be able to selectively pick up or drop off aplurality of processing units. In some embodiments, a fluid handlingdevice may be able to selectively aspirate and/or dispense a sampleusing one, two or more processing units. Any description of fluidhandling systems as described previously herein may apply to theprocessing units.

In one embodiment, a processing unit may be formed from molded plastic.The processing unit may be either commercially available or can be madeby injection molding with precise shapes and sizes. The units can becoated with capture reagents or other materials using method similar tothose used to coat microtiter plates but with the advantage that theycan be processed in bulk by placing them in a large vessel, addingcoating reagents and processing using sieves, holders, and the like torecover the pieces and wash them as needed. In some embodiments, thecapture reagents may be provided on an interior surface of theprocessing units.

A processing unit can offer a rigid support on which a reactant can beimmobilized. The processing unit may also be chosen to provideappropriate characteristics with respect to interactions with light. Forexample, the processing unit can be made of a material, such asfunctionalized glass, Si, Ge, GaAs, GaP, SiO₂, SiN₄, modified silicon,or any one of a wide variety of gels or polymers such as(poly)tetrafluoroethylene, (poly)vinylidenedifluoride, polystyrene,polycarbonate, polypropylene, Polymethylmethacylate (PMMA), ABS, orcombinations thereof. In an embodiment, a processing unit may comprisepolystyrene. Other appropriate materials may be used in accordance withthe present invention. Any of the materials described here, such asthose applying to tips and/or vessels may be used to form a processingunit. A transparent reaction site may be advantageous. In addition, inthe case where there is an optically transmissive window permittinglight to reach an optical detector, the surface may be advantageouslyopaque and/or preferentially light scattering. The processing unit mayoptionally be opaque and not permit the transmission of light therein.

A reactant may be immobilized at the capture surface of a processingunit. In some embodiments, the capture surface is provided on aninterior surface of the processing unit. In one example, the capturesurface may be provided in a lower portion of a processing tip orvessel.

The processing units can be dried following the last step ofincorporating a capture surface. For example, drying can be performed bypassive exposure to a dry atmosphere or via the use of a vacuum manifoldand/or application of clean dry air through a manifold.

In some embodiments, rather than using a capture surface on theprocessing unit, beads or other substrates may be provided to theprocessing units with capture surfaces provided thereon. One or morefree-flowing substrate may be provided with a capture surface. In someembodiments, the free-flowing substrate with a capture surface may beprovided within a fluid. In some embodiments, a bead may be magnetic.The bead may be coated with one or more reagents as known in the art. Amagnetic bead may be held at a desired location within the processingunit. The magnetic bead may be positioned using one or more magnet.

Beads may be useful for conducting one or more assay, including but notlimited to immunoassay, nucleic acid assay, or any of the other assaysdescribed elsewhere herein. The beads may be used during a reaction(e.g., chemical, physical, biological reaction). The beads may be usedduring one or more sample preparation step. The beads may be coated withone or more reagent. The beads themselves may be formed of reagents. Thebeads may be used for purification, mixing, filtering, or any otherprocesses. The beads may be formed of a transparent material,translucent material, and/or opaque material. The beads may be formed ofa thermally conductive or thermally insulative material. The beads maybe formed of an electrically conductive or electrically insulativematerial. The beads may accelerate a sample preparation and/or assaystep. The beads may provide an increased surface area that may reactwith one or more sample or fluid.

In alternate embodiments, beads or other solid materials may be providedto the assay units. The beads may be configured to dissolve undercertain conditions. For example, the beads may dissolve when in contactwith a fluid, or when in contact with an analyte or other reagents. Thebeads may dissolve at particular temperatures.

The beads may have any size or shape. The beads may be spherical. Thebeads may have a diameter of less than or equal to about 1 nm, 5 nm, 10nm, 50 nm, 100 nm, 200 nm, 300 nm, 500 nm, 750 nm, 1 μm, 2 μm, 3 μm, 5μm, 10 μm, 20 μm, 50 μm, 100 μm, 200 μm, 300 μm, 400 μm, 500 μm, 600 μm,700 μm, 800 μm, 900 μm, 1 mm, 1.2 mm, 1.5 mm, 2 mm, 2.5 mm, 3 mm, 4 mm,or 5 mm. The beads may be of the same size or differing sizes. The beadsmay include microparticles or nanoparticles.

A processing unit may have any dimension, including those describedelsewhere herein for tips and/or vessels. The processing unit may becapable of containing and/or confining a small volume of sample and/orother fluid, including volumes mentioned elsewhere herein.

A processing unit may be picked up and/or removed from a fluid handlingmechanism. For example, a processing tip or other processing unit may bepicked up by a pipette nozzle. The processing tip or other processingunit may be dropped off by a pipette nozzle. In some embodiments,processing units may be selectively individually picked up and/ordropped off. One or more group of processing units may be selectivelypicked up and/or dropped off. A processing unit may be picked up and/ordropped off using an automated mechanism. A processing unit may bepicked up and/or dropped off without requiring human intervention. Apipette may pick up and/or drop off a processing unit in accordance withdescriptions provided elsewhere herein.

A processing unit may be moved within a device and/or module using afluid handling mechanism. For example, a processing tip/vessel or otherprocessing unit may be transported using a pipette head. The processingtip/vessel or other processing unit may be transported in a horizontaldirection and/or vertical direction. The processing tip/vessel and/orprocessing unit may be transported in any direction. The processing unitmay be moved individually using the fluid handling mechanism. One ormore groups of processing units may be simultaneously moved using thefluid handling mechanism.

A processing unit may be shaped and/or sized to permit detection by adetection unit. The detection unit may be provided external to, inside,or integrated with the processing unit. In one example, the processingunit may be transparent. The processing unit may permit the detection ofan optical signal, audio signal, visible signal, electrical signal,magnetic signal, chemical signal, biological signal, motion,acceleration, weight, or any other signal by a detection unit.

A detector may be capable of detecting signals from individualprocessing units. The detector may differentiate signals received fromeach of the individual processing units. The detector may individuallytrack and/or follow signals from each of the individual processingunits. A detector may be capable of simultaneously detecting signalsfrom one or more groups of processing units. The detector may trackand/or follow signals from the one or more groups of processing units.

In some embodiments, magnetic particles or superparamagneticnanoparticles may be used in conjunction with vessels and miniaturizedmagnetic resonance to effect particular unit operations. Magneticparticles or superparamagnetic nanoparticles may be manipulated eithervia external magnetic fields, or via the pipette/fluid transfer device.Magnetic beads may be used for separations (when coated withantibodies/antigens/other capture molecules), for mixing (via agitationby external magnetic field), for concentrating analytes (either byselectively separating the analyte, or by separating impurities), etc.All these unit operations may be effectively carried out in smallvolumes with high efficiencies.

Reagent Unit

In accordance with an embodiment of the invention, an assay station, orany other portion of a module or device, may include one or more reagentunits. A reagent unit may be configured to contain and/or confine areagent that may be used in an assay. The reagent within the reagentunit may be used in a biological or chemical reaction. The reagent unitmay store one or more reagent prior to, during, or subsequent to areaction that may occur with the reagent. The biological and/or chemicalreactions may or may not take place external to the reagent units.

Reagents may include any of the reagents described in greater detailelsewhere herein. For example, reagents may include a sample diluent, adetector conjugate (for example, an enzyme-labeled antibody), a washsolution, and an enzyme substrate. Additional reagents can be providedas needed.

In some embodiments, a plurality of reagent units may be provided. Insome embodiments, one or more row of reagent units, and/or one or morecolumn of reagent units may be provided. In some embodiments, an m×narray of reagent units may be provided, wherein m, n are whole numbers.The reagent units may be provided in staggered rows or columns from eachother. In some embodiments, they may have any other configuration.

Any number of reagent units may be provided. For example there may bemore than and/or equal to about 1, 2, 3, 4, 5, 8, 10, 15, 20, 25, 30,40, 50, 60, 70, 80, 90, 100, 120, 150, 175, 200, 250, 300, 400, 500, or1000 reagent units.

Optionally, the same number of reagent units and assay units may beprovided. One or more reagent units may correspond to an assay unit. Oneor more assay units may correspond to a reagent unit. One or morereagent units may be movable relative to an assay unit. Alternative, oneor more assay unit may be movable relative to a reagent unit. An assayunit may be individually movable relative to a reagent unit.

Reagent units may be provided in a cartridge, card, or have any othersupporting structure. The reagent units may have the same orientation.For example reagent units may have one or more open end that may befacing in the same direction. Alternatively, reagent units may havedifferent orientations. In some examples, reagent units may be kept at avertical orientation. In other examples, reagent units may havehorizontal or vertical orientations, or any other angle of orientation.The reagent units may remain the same or may vary over time. Reagentunits may be provided on a supporting structure with assay units.Alternatively, reagent units may be provided on separate supportingstructures than assay units. Reagent units and assay units may besupported in separate portions of a supporting structure. Alternatively,they may be intermingled on a supporting structure.

The reagent units may be fluidically isolated or hydraulicallyindependent from one another. The reagent units may contain and/orconfine samples or other fluids that may be in fluid isolation from oneanother. The samples and/or other fluids contained within the reagentunits may be the same, or may vary from unit to unit. The system may becapable of tracking what each reagent unit contains. The system may becapable of tracking the location and history of each reagent unit.

The reagent units may be independently movable relative to one another,or another portion of the device or module. Thus, the fluids and/orsamples contained therein may be independently movable relative to oneanother or other portions of the device or module. A reagent unit may beindividually addressable. The location of each reagent unit may betracked. A reagent unit may be individually selected to receive and/orprovide a fluid. A reagent unit may be individually selected totransport a fluid. Fluid may be individually provided to or removed froma reagent unit. A reagent unit may be independently detectable.

Any description herein of individual reagent units may also apply togroups of reagent units. A group of reagent units may include one, two,or more reagent units. In some embodiments, reagent units within a groupmay be moved simultaneously. The location of groups of reagent units maybe tracked. Fluids may be simultaneously delivered and/or aspirated fromone or more group of reagent units. Detection may occur simultaneouslyto assay units within one or more groups of assay units.

The reagent units may have the form or characteristics of any of thetips or vessels as described elsewhere herein. For example, a reagentunit can be any of the tips or vessels described herein. Any descriptionherein of reagent units may also apply to tips or vessels, or anydescription of tips or vessels may also apply to the reagent units.

In some embodiments, a reagent unit may be a vessel. A reagent unit mayhave a first end and a second end. The first end and second end may beopposing one another. The first end and/or the second end may be open orclosed. In some embodiments, a first end may be open and a second endmay be closed. In alternate embodiments, the assay unit may have three,four, or more ends. The vessel may be covered by a septum and/or barrierto prevent evaporation and/or aerosolization to prevent reagent loss andcontamination of the device. The vessel may be disposable. Thiseliminates the requirement of externally filling reagents from a commonsource. This also allows better quality control and handling ofreagents. Additionally, this reduces contamination of the device and thesurroundings.

The reagent unit may have an interior surface and an exterior surface. Apassageway may connect the first and second ends of the reagent unit.The passageway may be a conduit or channel. The first and second ends ofthe assay tip may be in fluid communication with one another. Thediameter of the first end of the reagent unit may be greater than thediameter of the second end of the reagent unit. In some embodiments, theouter diameter of the first end of the reagent unit may be greater thanthe outer diameter of the second end of the reagent unit. Alternatively,the diameters may be the same, or the outer diameter of the second endmay be greater than the outer diameter of the first end. An innerdiameter of the first end of the reagent unit may be greater than theinner diameter of the second end of the reagent unit. Alternatively, adiameter and/or inner diameter of the reagent unit may be the same atthe first and second ends. In some embodiments, the second end may beheld below the first end of the reagent unit. Alternatively the relativepositions of the first and second ends may vary. An open end of thereagent unit may be oriented upwards, or may be held higher than aclosed end.

In some embodiments, a reagent unit may have a cap or closure. The capor closure may be capable of blocking an open end of the reagent unit.The cap or closure may be selectively applied to close or open the openend of the reagent unit. The cap or closure may have one or moreconfiguration as illustrated elsewhere herein or as known in the art.The cap or closure may form an airtight seal that may separate thecontents of the reagent unit from the ambient environment.

As previously described regarding tips and/or vessels, a reagent unitmay be picked up using a fluid handling device. For example, a pipetteor other fluid handling device may connect to the reagent unit. Apipette nozzle or orifice may interface with an end of the reagent unit.In some embodiments, a fluid-tight seal may be formed between the fluidhandling device and the reagent unit. A reagent unit may be attached toand/or detached from the fluid handling device. The fluid handlingdevice may move the reagent unit from one location to another.Alternatively, the reagent unit is not connected to the fluid handlingdevice. Any other automated device or process may be used to move ormanipulate an assay unit. A reagent unit may be moved or manipulatedwithout the intervention of a human.

A reagent unit may be configured to accept an assay unit. In someembodiments, a reagent unit may include an open end through which atleast a portion of an assay unit may be inserted. In some embodiments,the assay unit may be entirely inserted within the reagent unit. An openend of the reagent unit may have a greater diameter than at least one ofthe open ends of the assay unit. In some instances, an inner diameter ofan open end of the reagent unit may be greater than an outer diameter ofat least one of the open ends of the assay unit. In some embodiments, areagent unit may be shaped or may include one or more feature that maypermit the assay unit to be inserted a desired amount within the reagentunit. The assay unit may or may not be capable of being insertedcompletely into the reagent unit.

An assay unit may dispense to and/or aspirate a fluid from the reagentunit. A reagent unit may provide a fluid, such as a reagent, that may bepicked up by the assay unit. The assay unit may optionally provide afluid to the reagent unit. Fluid may be transferred through the open endof a reagent unit and an open end of the assay unit. The open ends ofthe assay unit and the reagent unit may permit the interior portions ofthe assay unit and the reagent unit to be brought into fluidcommunication with one another. In some embodiments, an assay unit maybe located above the reagent unit during said dispensing and/oraspiration.

Alternatively, fluid transfer between the reagent unit and the assayunit may be done by a fluid handling device. One or several such fluidtransfers might happen simultaneously. The fluid handling device in oneembodiment might be a pipette.

In one example, a reagent for a chemical reaction may be provided withina reagent unit. An assay unit may be brought into the reagent unit andmay aspirate the reagent from the reagent unit. A chemical reaction mayoccur within the assay unit. The excess fluid from the reaction may bedispensed from the assay unit. The assay unit may pick up a washsolution. The wash solution may be expelled from the assay unit. Thewashing step may occur one, two, three, four, five, or more times. Thewash solution may optionally be picked up and/or dispensed to a reagentunit. This may reduce background signal interference. A detector maydetect one or more signal from the assay unit. The reduced backgroundsignal interference may permit increased sensitivity of signals detectedfrom the assay unit. An assay tip format may be employed, which mayadvantageously provide easy expulsion of fluids for improved washingconditions.

A fluid handling device or any other automated device may be able topick up or drop off an individual assay unit. A fluid handling device orother automated device may be able to simultaneously pick up or drop offa plurality of assay units. A fluid handling device or other automateddevice may be able to selectively pick up or drop off a plurality ofassay units. In some embodiments, a fluid handling device may be able toselectively aspirate and/or dispense a sample using one, two or moreassay units. Any description of fluid handling systems as describedpreviously herein may apply to the assay units.

In one embodiment, a reagent unit may be formed from molded plastic. Thereagent unit may be either commercially available or can be made byinjection molding with precise shapes and sizes. The units can be coatedwith capture reagents using method similar to those used to coatmicrotiter plates but with the advantage that they can be processed inbulk by placing them in a large vessel, adding coating reagents andprocessing using sieves, holders, and the like to recover the pieces andwash them as needed. In some embodiments, the capture reagents may beprovided on an interior surface of the reagent units. Alternativelyreagent units may be uncoated, or may be coated with other substances.

A reagent unit can offer a rigid support. The reagent unit may be chosento provide appropriate characteristics with respect to interactions withlight. For example, the reagent unit can be made of a material, such asfunctionalized glass, Si, Ge, GaAs, GaP, SiO₂, SiN₄, modified silicon,or any one of a wide variety of gels or polymers such as(poly)tetrafluoroethylene, (poly)vinylidenedifluoride, polystyrene,polycarbonate, polypropylene, PMMA, ABS, or combinations thereof. In anembodiment, an assay unit may comprise polystyrene. Other appropriatematerials may be used in accordance with the present invention. Any ofthe materials described here, such as those applying to tips and/orvessels may be used to form a reagent unit. A transparent reaction sitemay be advantageous. In addition, in the case where there is anoptically transmissive window permitting light to reach an opticaldetector, the surface may be advantageously opaque and/or preferentiallylight scattering.

A reagent unit may or may not offer a capture surface, such as thosedescribed for assay units. Similarly, a reagent unit may or may notemploy beads or other substrates to provide capture surfaces. Anydescription relating to beads or other capture surfaces for assay unitsor processing units may also optionally be applied to reagent units.

A reagent unit may or may not have a reaction site. Any descriptionherein of a reaction site for an assay unit may also apply to a reagentunit.

A reagent unit may have any dimension, including those describedelsewhere herein for tips and/or vessels. The reagent unit may becapable of containing and/or confining a small volume of sample and/orother fluid, including volumes mentioned elsewhere herein.

A reagent unit may be stationary within a device and/or module.Alternatively, a reagent unit may be movable relative to the deviceand/or module. A reagent unit may be picked up and/or moved using afluid handling mechanism or any other automated process. For example, areagent unit may be picked up by a pipette nozzle, such as in a mannerdescribed elsewhere for an assay unit.

Relative movement may occur between the assay unit and the reagent unit.The assay unit and/or reagent unit may move relative to one another.Assay units may move relative to one another. Reagent units may moverelative to one another. Assay units and/or reagent units may beindividually movable relative to the device and/or module.

A reagent unit may be shaped and/or sized to permit detection by adetection unit. The detection unit may be provided external to, inside,or integrated with the reagent unit. In one example, the reagent unitmay be transparent. The reagent unit may permit the detection of anoptical signal, audio signal, visible signal, electrical signal,magnetic signal, motion, acceleration, weight, or any other signal by adetection unit.

A detector may be capable of detecting signals from individual reagentunits. The detector may differentiate signals received from each of theindividual reagent units. The detector may individually track and/orfollow signals from each of the individual reagent units. A detector maybe capable of simultaneously detecting signals from one or more groupsof reagent units. The detector may track and/or follow signals from theone or more groups of reagent units. Alternatively, the detector neednot detect signals from individual reagents. In some embodiments thedevice and/or system may keep track of the identity of reagents or otherfluids provided within the reagent units, or information associated withthe reagents or other fluids.

As previously mentioned reagent units may include one or more reagentstherein. Reagents may include a wash buffer, enzyme substrate, dilutionbuffer, or conjugates (such as enzyme labeled conjugates). Examples ofenzyme labeled conjugates may include polyclonal antibodies, monoclonalantibodies, or may be labeled with enzyme that can yield a detectablesignal upon reaction with an appropriate substrate. Reagents may alsoinclude DNA amplifiers, sample diluents, wash solutions, samplepre-treatment reagents (including additives such as detergents),polymers, chelating agents, albumin-binding reagents, enzyme inhibitors,enzymes (e.g., alkaline phosphatase, horseradish peroxide),anticoagulants, red-cell agglutinating agents, or antibodies. Any otherexamples of reagents described elsewhere herein may also be containedand/or confined within a reagent unit.

Dilution

The device and/or module may permit the use of one or more diluents inaccordance with an embodiment of the invention. Diluent may be containedin one or more reagent unit, or any other unit that may contain and/orconfine the diluents. The diluents may be provided in a tip, vessel,chamber, container, channel, tube, reservoir, or any other component ofthe device and/or module. Diluent may be stored in a fluidicallyisolated or hydraulically independent component. The fluidicallyisolated or hydraulically independent component may be stationary or maybe configured to move relative to one or more portion of the deviceand/or module.

In some embodiments, diluents may be stored in diluents units, which mayhave any characteristics of reagent units as described elsewhere herein.The diluents units may be stored in the same location as the rest of thereagent units, or may be stored remotely relative to the rest of thereagent units.

Any examples of diluents known in the art may be employed. Diluent maybe capable of diluting or thinning a sample. In most instances, thediluents do not cause a chemical reaction to occur with the sample. Adevice may employ one type of diluents. Alternatively, the device mayhave available or employ multiple types of diluents. The system may becapable of tracking diluents and/or various types of diluents. Thus, thesystem may be capable of accessing a desired type of diluents. Forexample, a tip may pick up a desired diluent.

In some embodiments, diluents may be provided to a sample. The diluentsmay dilute the sample. The sample may become less concentrated with theaddition of a diluent. The degree of dilution may be controlledaccording to one or more protocol or instructions. In some instances,the protocol or instructions may be provided from an external device,such as a server. Alternatively, the protocol or instructions may beprovided on-board the device or cartridge or vessel. Thus, a serverand/or the device may be capable of variable dilution control. Bycontrolling the degree of dilution, the system may be capable ofdetecting the presence or concentration of one or more analytes that mayvary over a wide range. For example, a sample may have a first analytehaving a concentration that would be detectable over a first range, anda second analyte having a concentration that would be detectable overthe second range. The sample may be divided and may or may not havevarying amounts of diluents applied to bring the portions of the sampleinto a detectable range for the first and second analytes. Similarly, asample may or may not undergo varying degrees of enrichment to bringanalytes to a desired concentration for detection.

Dilution and/or enrichment may permit the one, two, three or moreanalytes having a wide range of concentrations to be detected. Forexamples, analytes differing by one or more, two or more, three or more,four or more, five or more, six or more, seven or more, eight or more,nine or more, or ten or more degrees of magnitude may be detected from asample.

In some embodiments, a sample may be combined with diluents in an assaytip or other type of tip described elsewhere herein. An assay tip mayaspirate a diluent. The assay tip may pick up the diluents from areagent unit. The diluents may or may not be combined with the samplewithin the assay tip.

In another example, a diluents and/or sample may be combined in areagent unit or other types of vessels described elsewhere herein. Forexample, a diluents may be added to a sample in a reagent unit, or asample may be added to a diluents in the reagent unit.

In some embodiments, one or more mixing mechanism may be provided.Alternatively, no separate mixing mechanism is needed. The assay unit,reagent unit, or any other tip, vessel, or compartment combining asample and diluents may be capable of moving, thereby effecting amixing.

Varying amounts of diluents and/or samples may be combined to achieve adesired level of dilution. Protocols may determine the relativeproportion of diluents and sample to combine. In some embodiments, theportion of sample to diluent may be less than and/or equal to about1:1,000,000, 1:100,000, 1:10,000, 1:1,000, 1:500, 1:100, 1:50, 1:10,1:5, 1:3, 1:2, 1:1, or greater than and/or equal to 2:1, 3:1, 5:1, 10:1,50:1, 100:1, 500:1, 1,000:1, 10,000:1, 100,000:1, or 1,000,000:1. Thediluted sample may be picked up from the reagent unit using an assaytip, where one or more chemical reaction may occur.

A desired amount of diluents may be provided in accordance with one ormore set of instructions. In some embodiments, the amount of dilutionprovided may be controlled by a fluid handling system. For example, anassay tip may pick up a desired amount of diluents and dispense it to adesired location. The volume of diluents picked up by the assay tip maybe controlled with a high degree of sensitivity. For example, the amountof diluents picked up may have any of the volumes of fluids or samplesdiscussed elsewhere herein. In some embodiments, an assay tip may pickup a desired amount of diluents in one turn. Alternatively, an assay tipmay pick up and dispense diluents multiple times in order to achieve adesired degree of dilution.

Dilution of a sample may occur during a sample pre-treatment step. Asample may be diluted prior to undergoing a chemical reaction.Alternatively, dilution may occur during a chemical reaction and/orsubsequent to a chemical reaction.

The dilution factor may be optimized in real-time for each assaydepending on the assay requirements. In one embodiment, real-timedetermination of a dilution scheme can be performed by knowledge of allassays to be performed. This optimization may take advantage of multipleassays using identical dilution. The aforementioned dilution scheme mayresult in higher precision of final diluted sample.

Dilution of a sample may be performed serially or in a single step. Fora single-step dilution, a selected quantity of sample may be mixed witha selected quantity of diluent, in order to achieve a desired dilutionof the sample. For a serial dilution, two or more separate sequentialdilutions of the sample may be performed in order to achieve a desireddilution of the sample. For example, a first dilution of the sample maybe performed, and a portion of that first dilution may be used as theinput material for a second dilution, to yield a sample at a selecteddilution level.

For dilutions described herein, an “original sample” refers to thesample that is used at the start of a given dilution process. Thus,while an “original sample” may be a sample that is directly obtainedfrom a subject, it may also include any other sample (e.g. sample thathas been processed or previously diluted in a separate dilutionprocedure) that is used as the starting material for a given dilutionprocedure.

In some embodiments, a serial dilution of a sample may be performed witha device described herein as follows. A selected quantity (e.g. volume)of an original sample may be mixed with a selected quantity of diluent,to yield a first dilution sample. The first dilution sample (and anysubsequent dilution samples) will have: i) a sample dilution factor(e.g. the amount by which the original sample is diluted in the firstdilution sample) and ii) an initial quantity (e.g. the total quantity ofthe first dilution sample present after combining the selected quantityof original sample and selected quantity of diluent). For example, 10microliters of an original sample may be mixed with 40 microliters ofdiluent, to yield a first dilution sample having a 5-fold dilutionfactor and an initial quantity of 50 microliters. Next, a selectedquantity of the first dilution sample may be mixed with a selectedquantity of diluent, to yield a second dilution sample. For example, 5microliters of the first dilution sample may be mixed with 95microliters of diluent, to yield a second dilution sample having an100-fold dilution factor and an initial quantity of 100 microliters. Foreach of the above dilution steps, the original sample, dilutionsample(s), and diluent may be stored or mixed in fluidically isolatedvessels. Sequential dilutions may continue in the preceding manner foras many steps as needed to reach a selected sample dilutionlevel/dilution factor.

In devices provided herein, an original sample may be diluted, forexample, at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40,50, 75, 100, 200, 300, 400, 500, 1,000, 5,000, 10,000, 20,000, 50,000,or 100,000-fold, by either a single-step or serial dilution procedure.In some embodiments, a single original sample may be diluted to reachmultiple different selected sample dilution factors (e.g. a singleoriginal sample may be diluted to generate samples which are diluted5-fold, 10-fold, 25-fold, 100-fold, 200-fold, 500-fold, and 1000-fold).In some embodiments, a device may be configured to perform a 2, 3, 4, 5,6, 7, 8, 9, 10, or more step serial dilution. A device may be configuredto dilute 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more different originalsamples within the same device (e.g. a device may dilute bothEDTA-containing and heparin-containing plasma samples at the same time).In some embodiments, a device provided herein contains a controllerwhich is configured to instruct a sample handling system within thedevice to perform one or more sample handling steps to prepare any ofthe dilutions of sample described above or elsewhere herein. Thecontroller may direct the device to use 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,or more different diluents for different dilution procedures. Thecontroller may contain a protocol for performing the dilutions. Theprotocol may be stored or generated on-the-fly. The protocol may be sentfrom an external device to the sample processing device, or stored orgenerated on the sample processing device.

In some embodiments, one or more steps of a dilution procedure may beperformed with a sample handling system. The sample handling system maybe a pipette or other fluid handling apparatus. The sample handlingsystem may be configured for obtaining a selected quantity of a sampleor diluent from a fluidically isolated vessel containing the sample ordiluent, and transporting the selected quantity of sample or diluent toa different fluidically isolated vessel. During the dilution of asample, the diluent may be deposited in a vessel before the sample isadded to the diluent. Alternatively, the sample may be deposited in avessel before the diluent is added to the sample. In other embodiments,the sample and diluent may be in the same fluid circuit.

Dilution of samples may facilitate the performance of a large number ofassays with a small amount of original sample. In some situations,dilution of an original sample into multiple dilution samples havingdifferent dilution factors may, for example: i) reduce waste of sample,for example, by only using the minimum amount of original samplerequired to perform each assay (i.e. by not using samples that are moreconcentrated than necessary to perform the assay); ii) increase thetotal number of assays that may be performed with a given amount oforiginal sample, for example, by the reduction of waste of sample; andiii) increase the variety of assays that may be performed with anoriginal sample, for example, by dilution of the original sample todifferent sample dilution factors, where different sample dilutionfactors are needed to perform different assays (for example, if oneassay requires a high sample concentration in order to efficientlydetect an analyte that is not abundant in the sample, and if anotherassay requires a low sample concentration in order to efficiently detectan analyte that is abundant in the sample).

Washing

The device and/or module may permit washing in accordance with anembodiment of the invention. A wash solution may be contained in one ormore reagent unit, or any other unit that may contain and/or confine thewash solution. The wash solution may be provided in a tip, vessel,chamber, container, channel, tube, reservoir, or any other component ofthe device and/or module. A wash solution may be stored in a fluidicallyisolated or hydraulically independent component. The fluidicallyisolated or hydraulically independent component may be stationary or maybe configured to move relative to one or more portion of the deviceand/or module.

In some embodiments, wash solution may be stored in wash units, whichmay have any characteristics of reagent units as described elsewhereherein. The wash units may be stored in the same location as the rest ofthe reagent units, or may be stored remotely relative to the rest of thereagent units.

Any examples of wash solutions known in the art may be employed. Washsolutions may be capable of removing unbound and/or unreacted reactants.For examples, a chemical reaction may occur between a sample containingan analyte and an immobilized reactant, that may cause an analyte tobind to a surface. The unbound analytes may be washed away. In someembodiments, a reaction may cause the emission of an optical signal,light, or any other sort of signal. If unreacted reactants remain in theproximity, they may cause interfering background signal. It may bedesirable to remove the unreacted reactants to reduce interferingbackground signal and permit the reading of the bound analytes. In someinstances, the wash solution does not cause a chemical reaction to occurbetween the wash solution and the sample.

A device may employ one type of wash solutions. Alternatively, thedevice may have available or employ multiple types of wash solutions.The system may be capable of tracking wash solutions and/or varioustypes of wash solutions. Thus, the system may be capable of accessing adesired type of wash solution. For example, a tip may pick up a desiredwash solution.

In some embodiments, a wash solution may be provided to a sample. Thewash solution may dilute the sample. The sample may become lessconcentrated with the addition of a wash solution. The degree of washingmay be controlled according to one or more protocol or instructions. Bycontrolling the degree of washing, the system may be capable ofdetecting the presence or concentration of one or more analytes with adesired sensitivity. For example, increased amounts of washing mayremove undesirable reagents or sample that may cause interferingbackground noise.

In some embodiments, a wash solution may be provided to an assay tip orother type of tip described elsewhere herein. An assay tip may aspiratea wash solution. The assay tip may pick up the wash solutions from awash unit. The wash solution may or may not be dispensed back outthrough the assay tip. The same opening of an assay tip may bothaspirate and dispense the wash solution. For example, an assay tip mayhave a bottom opening that may be used to both pick up and expel a washsolution. The assay tip may have both a bottom opening and a topopening, where the bottom opening may have a smaller diameter than thetop opening. Expelling the wash solution through the bottom opening maypermit more effective expulsion of the wash solution than if the bottomof the assay tip were closed.

In another example, a wash solution and/or sample may be combined in areagent unit or other types of vessels described elsewhere herein. Forexample, a wash solution may be added to a sample in a reagent unit, ora sample may be added to a wash solution in the reagent unit. The washsolution may be expelled in any manner. In some embodiments, acombination of the wash solution and/or sample may be picked up by anassay tip.

A desired amount of wash solution may be provided in accordance with oneor more set of instructions. In some embodiments, the amount of washsolution provided may be controlled by a fluid handling system. Forexample, an assay tip may pick up a desired amount of wash solution anddispense it. The volume of wash solution picked up by the assay tip maybe controlled with a high degree of sensitivity. For example, the amountof wash solution picked up may have any of the volumes of fluids orsamples discussed elsewhere herein. In some embodiments, an assay tipmay pick up a desired amount of wash solution in one turn.Alternatively, an assay tip may pick up and dispense wash solutionmultiple times in order to achieve a desired degree of washing.

Varying numbers of wash cycles may occur to provide a desiredsensitivity of detection. Protocols may determine the number of washcycles. For example, greater than, and/or equal to about one, two,three, four, five, six, seven, eight, nine, ten, eleven, or twelve washcycles may occur. The wash solution may be picked up from the wash unitusing an assay tip, and may be expelled from the assay tip.

Washing may occur subsequent to undergoing a chemical reaction.Alternatively, washing may occur during a chemical reaction and/or priorto a chemical reaction.

Contamination Reduction

The device and/or module may permit contamination prevention and/orreduction in accordance with an embodiment of the invention. Forexample, a touch-off pad may be provided. The touch-off pad may beformed of an absorbent material. For example, the touch-off pad may be asponge, textile, gel, porous material, capillary or have any featurethat may absorb or wick away a fluid that may come into contact with thepad. An assay tip may be brought into contact with the touch-off pad,which may result in fluid from the assay tip in proximity to thetouch-off pad being absorbed by the pad. In some embodiments, an assaytip may be brought to a touch-off pad in a manner such that the assaytip does not contact a portion of the pad that has previously beencontacted. In some instances, liquid is not placed in the same place asa liquid has been previously touched off. The assay tips may be broughtto the pad in a way so that the contact points are spaced apart so thata different contact point is used whenever an assay tip touches the pad.One or more controller may determine the location of the touch-off padthat an assay tip may contact next. The controller may keep track ofwhat points on the pad have already been contacted by an assay tip. Theassay pad may be absorbent.

The assay tip may be wiped by the pad. The excess fluid or undesiredfluid from the assay tip may be removed from the assay tip. For example,an open end, such as a bottom end, of the assay tip may be brought intocontact with the touch-off pad. The pad may be formed from an absorbentmaterial that may wick the fluid away from the assay tip. Thus, as anassay tip, or other component of the device, may move throughout amodule and/or device, the likelihood of excess fluid or undesired fluidfrom contaminating other portions of the module and/or device may bereduced. In one non-limiting example, an absorbent pad is part of thecartridge and it is configured to wick fluid away from tips, reducingcarry over. In some embodiments, an absorbant pad may be any location ina device accessible by a sample handling system. Use of an absorbent padwith pipetting or other tip-related liquid transfer methods may increasethe accuracy and precision of the fluid transfer and may lower thecoefficient of variation of transferring fluid with the liquid transfermethods.

Another example of a contamination prevention and/or reduction mechanismmay include applying a coating or covering to an assay tip or othercomponent of the device. For example, an assay tip may be brought intocontact with a melted wax, oil (such as mineral oil), or a gel. In someembodiments, the wax, oil, or gel may harden. Hardening may occur as thematerial cools and/or is exposed to air. Alternatively, they need notharden. The coating surface, such as a wax, oil, or gel, may besufficiently viscous to remain on the assay tip or other component ofthe device. In one example, an open end of the assay tip may be broughtinto contact with the coating material, which may cover the open end ofthe assay tip, sealing the contents of the assay tip.

Additional examples of contamination prevention and/or reduction may bea waste chamber to accept used assay tips, a component that may put oneor more cap on used portions of assay tips, a heater or fan, orultraviolet light emitted onto one or more components or subsystems, orany other component that may reduce the likelihood of contamination anyother component that may reduce the likelihood of contamination. In someembodiments, the fluid handling components of the device do not requireregular decontamination as the fixed components of the device do notnormally come in direct contact with the sample. The fluid handlingdevice may be capable of periodical self-sanitization, such as byaspirating cleaning agents (e.g., ethanol) from a tank using thepipette. The fluid handling apparatus, and other device resources, canalso be decontaminated, sterilized, or disinfected by a variety of othermethods, including UV irradiation.

Filter

The device and/or modules may include other components, which may permitone or more function as described elsewhere herein. For example, thedevice and/or module may have a filter that may permit the separation ofa sample by particle size, density, or any other feature. For example, aparticle or fluid having a particle size smaller than a threshold sizemay pass through a filter while other particles having a size greaterthan the threshold size do not. In some embodiments, a plurality offilters may be provided. The plurality of filters may have the same sizeor different sizes, which may permit sorting of different sizes ofparticles into any number of groups.

Centrifuge

In accordance with some embodiments of the invention, a system mayinclude one or more centrifuge. A device may include one or morecentrifuge therein. For example, one or more centrifuge may be providedwithin a device housing. A module may have one or more centrifuge. One,two, or more modules of a device may have a centrifuge therein. Thecentrifuge may be supported by a module support structure, or may becontained within a module housing. The centrifuge may have a form factorthat is compact, flat and requires only a small footprint. In someembodiments, the centrifuge may be miniaturized for point-of-serviceapplications but remain capable of rotating at high rates, equal to orexceeding about 10,000 rpm, and be capable of withstanding g-forces ofup to about 1200 m/s² or more.

A centrifuge may be configured to accept one or more sample. Acentrifuge may be used for separating and/or purifying materials ofdiffering densities. Examples of such materials may include viruses,bacteria, cells, proteins, environmental compositions, or othercompositions. A centrifuge may be used to concentrate cells and/orparticles for subsequent measurement.

A centrifuge may have one or more cavity that may be configured toaccept a sample. The cavity may be configured to accept the sampledirectly within the cavity, so that the sample may contact the cavitywall. Alternatively, the cavity may be configured to accept a samplevessel that may contain the sample therein. Any description herein ofcavity may be applied to any configuration that may accept and/orcontain a sample or sample container. For example, cavities may includeindentations within a material, bucket formats, protrusions with hollowinteriors, members configured to interconnect with a sample container.Any description of cavity may also include configurations that may ormay not have a concave or interior surface. Examples of sample vesselsmay include any of the vessel or tip designs described elsewhere herein.Sample vessels may have an interior surface and an exterior surface. Asample vessel may have at least one open end configured to accept thesample. The open end may be closeable or sealable. The sample vessel mayhave a closed end. The sample vessel may be a nozzle of the fluidhandling apparatus, which apparatus may act as a centrifuge to spin afluid in the nozzle, the tip or another vessel attached to such anozzle.

A centrifuge may have one or more, two or more, three or more, four ormore, five or more, six or more, eight or more, 10 or more, 12 or more,15 or more, 20 or more, 30 or more, or 50 or more cavities configured toaccept a sample or sample vessel.

In some embodiments, the centrifuge may be configured to accept a smallvolume of sample. In some embodiments, the cavity and/or sample vesselmay be configured to accept a sample volume of 1,000 μL or less, 500 μLor less, 250 μL or less, 200 μL or less, 175 μL or less, 150 μL or less,100 μL or less, 80 μL or less, 70 μL or less, 60 μL or less, 50 μL orless, 30 μL or less, 20 μL or less, 15 μL or less, 10 μL or less, 8 μLor less, 5 μL or less, 1 μL or less, 500 nL or less, 300 nL or less, 100nL or less, 50 nL or less, 10 nL or less, 1 nL or less, 500 pL or less,100 pL or less 50 pL or less, 10 pL or less 5 pL or less, or 1 pL orless. In some embodiments, centrifuge may be configured such that thetotal volume that the centrifuge is configured to accept (e.g. thecombined volume that may be accepted by the total of all the cavitiesand/or sample vessels in the centrifuge) is 10 ml or less, 5 ml or less,4 ml or less, 3 ml or less, 2 ml or less, 1 ml or less, 750 μl or less,500 μl or less, 400 μl or less, 300 μl or less, 200 μl or less, 100 μlor less, 50 μl or less, 40 μl or less, 30 μl or less, 20 μl or less, 10μl or less, 8 μl or less, 6 μl or less, 4 μl or less, or 2 μl or less.In some embodiments, the centrifuge may contain 50 or less, 40 or less,30 or less, 29 or less, 28 or less, 27 or less, 26 or less, 25 or less,24 or less, 23 or less, 22 or less, 21 or less, 20 or less, 19 or less,18 or less, 17 or less, 16 or less, 15 or less, 14 or less, 13 or less,12 or less, 11 or less, 10 or less, 9 or less, 8 or less, 7 or less, 6or less, 5 or less, 4 or less, 3 or less, 2 or less, or 1 cavitiesand/or sample vessels, which are configured to accept, in total, avolume of 10 ml or less, 5 ml or less, 4 ml or less, 3 ml or less, 2 mlor less, 1 ml or less, 750 μl or less, 500 μl or less, 400 μl or less,300 μl or less, 200 μl or less, 100 μl or less, 50 μl or less, 40 μl orless, 30 μl or less, 20 μl or less, 10 μl or less, 8 μl or less, 6 μl orless, 4 μl or less, or 2 μl or less.

In some embodiments, the centrifuge may have a cover that may containthe sample within the centrifuge. The cover may prevent the sample foraerosolizing and/or evaporating. The centrifuge may optionally have afilm, oil (e.g., mineral oil), wax, or gel that may contain the samplewithin the centrifuge and/or prevent it from aerosolizing and/orevaporating. The film, oil, wax, or gel may be provided as a layer overa sample that may be contained within a cavity and/or sample vessel ofthe centrifuge.

A centrifuge may be configured to rotate about an axis of rotation. Acentrifuge may be able to spin at any number of rotations per minute.For example, a centrifuge may spin up to a rate of 100 rpm, 1,000 rpm,2,000 rpm, 3,000 rpm, 5,000 rpm, 7,000 rpm, 10,000 rpm, 12,000 rpm,15,000 rpm, 17,000 rpm, 20,000 rpm, 25,000 rpm, 30,000 rpm, 40,000 rpm,50,000 rpm, 70,000 rpm, or 100,000 rpm. At some points in time, acentrifuge may remain at rest, while at other points in time, thecentrifuge may rotate. A centrifuge at rest is not rotating. Acentrifuge may be configured to rotate at variable rates. In someembodiments, the centrifuge may be controlled to rotate at a desirablerate. In some embodiments, the rate of change of rotation speed may bevariable and/or controllable.

In some embodiments, the axis of rotation may be vertical.Alternatively, the axis of rotation may be horizontal, or may have anyangle between vertical and horizontal (e.g., about 15, 30, 45, 60, or 75degrees). In some embodiments, the axis of rotation may be in a fixeddirection. Alternatively, the axis of rotation may vary during the useof a device. The axis of rotation angle may or may not vary while thecentrifuge is rotating.

A centrifuge may comprise a base. The base may have a top surface and abottom surface. The base may be configured to rotate about the axis ofrotation. The axis of rotation may be orthogonal to the top and/orbottom surface of the base. In some embodiments, the top and/or bottomsurface of the base may be flat or curved. The top and bottom surfacemay or may not be substantially parallel to one another.

In some embodiments, the base may have a circular shape. The base mayhave any other shape including, but not limited to, an elliptical shape,triangular shape, quadrilateral shape, pentagonal shape, hexagonalshape, or octagonal shape.

The base may have a height and one or more lateral dimension (e.g.,diameter, width, or length). The height of the base may be parallel tothe axis of rotation. The lateral dimension may be perpendicular to theaxis of rotation. The lateral dimension of the base may be greater thanthe height. The lateral dimension of the base may be 2 times or more, 3times or more, 4 times or more, 5 times or more, 6 times or more, 8times or more, 10 times or more, 15 times or more, or 20 times or moregreater than the height.

The centrifuge may have any size. For example, the centrifuge may have afootprint of about 200 cm² or less, 150 cm² or less, 100 cm² or less, 90cm² or less, 80 cm² or less, 70 cm² or less, 60 cm² or less, 50 cm² orless, 40 cm² or less, 30 cm² or less, 20 cm² or less, 10 cm² or less, 5cm² or less, or 1 cm² or less. The centrifuge may have a height of about5 cm or less, 4 cm or less, 3 cm or less, 2.5 cm or less, 2 cm or less,1.75 cm or less, 1.5 cm or less, 1 cm or less, 0.75 cm or less, 0.5 cmor less, or 0.1 cm or less. In some embodiments, the greatest dimensionof the centrifuge may be about 15 cm or less, 10 cm or less, 9 cm orless, 8 cm or less, 7 cm or less, 6 cm or less, 5 cm or less, 4 cm orless, 3 cm or less, 2 cm or less, or 1 cm or less.

The centrifuge base may be configured to accept a drive mechanism. Adrive mechanism may be a motor, or any other mechanism that may enablethe centrifuge to rotate about an axis of rotation. The drive mechanismmay be a brushless motor, which may include a brushless motor rotor anda brushless motor stator. The brushless motor may be an induction motor.The brushless motor rotor may surround the brushless motor stator. Therotor may be configured to rotate about a stator about an axis ofrotation.

The base may be connected to or may incorporate the brushless motorrotor, which may cause the base to rotate about the stator. The base maybe affixed to the rotor or may be integrally formed with the rotor. Thebase may rotate about the stator and a plane orthogonal to the axis ofrotation of the motor may be coplanar with a plane orthogonal to theaxis of rotation of the base. For example, the base may have a planeorthogonal to the base axis of rotation that passes substantiallybetween the upper and lower surface of the base. The motor may have aplane orthogonal to the motor axis of rotation that passes substantiallythrough the center of the motor. The base planes and motor planes may besubstantially coplanar. The motor plane may pass between the upper andlower surface of the base.

A brushless motor assembly may include the rotor and stator. The motorassembly may include the electronic components. The integration of abrushless motor into the rotor assembly may reduce the overall size ofthe centrifuge assembly. In some embodiments, the motor assembly doesnot extend beyond the base height. In other embodiments, the height ofthe motor assembly is no greater than 1.5 times the height of the base,than twice the height of the base, than 2.5 times the height of thebase, than three times the height of the base, than four times theheight of the base, or five times the height of the base. The rotor maybe surrounded by the base such that the rotor is not exposed outside thebase.

The motor assembly may effect the rotation of the centrifuge withoutrequiring a spindle/shaft assembly. The rotor may surround the statorwhich may be electrically connected to a controller and/or power source.

In some embodiments, the cavity may be configured to have a firstorientation when the base is at rest, and a second orientation when thebase is rotating. The first orientation may be a vertical orientationand a second orientation may be a horizontal orientation. The cavity mayhave any orientation, where the cavity may be more than and/or equal toabout 0 degrees, 5 degrees, 10 degrees, 15 degrees, 20 degrees, 25degrees, 30 degrees, 35 degrees, 40 degrees, 45 degrees, 50 degrees, 55degrees, 60 degrees, 65 degrees, 70 degrees, 75 degrees, 80 degrees, 85degrees, or 90 degrees from vertical and/or the axis of rotation. Insome embodiments, the first orientation may be closer to vertical thanthe second orientation. The first orientation may be closer to parallelto the axis of rotation than the second orientation. Alternatively, thecavity may have the same orientation regardless of whether the base isat rest or rotating. The orientation of the cavity may or may not dependon the speed at which the base is rotating.

The centrifuge may be configured to accept a sample vessel, and may beconfigured to have the sample vessel at a first orientation when thebase is at rest, and have the sample vessel at a second orientation whenthe base is rotating. The first orientation may be a verticalorientation and a second orientation may be a horizontal orientation.The sample vessel may have any orientation, where the sample vessel maybe more than and/or equal to about 0 degrees, 5 degrees, 10 degrees, 15degrees, 20 degrees, 25 degrees, 30 degrees, 35 degrees, 40 degrees, 45degrees, 50 degrees, 55 degrees, 60 degrees, 65 degrees, 70 degrees, 75degrees, 80 degrees, 85 degrees, or 90 degrees from vertical. In someembodiments, the first orientation may be closer to vertical than thesecond orientation. Alternatively, the sample vessel may have the sameorientation regardless of whether the base is at rest or rotating. Theorientation of the vessel may or may not depend on the speed at whichthe base is rotating.

FIG. 36 shows an example of a centrifuge provided in accordance with anembodiment of the invention. The centrifuge may include a base 3600having a bottom surface 3602 and/or top surface 3604. The base maycomprise one, two or more wings 3610 a, 3610 b.

A wing may be configured to fold over an axis extending through thebase. In some embodiments, the axis may form a secant through the base.An axis extending through the base may be a foldover axis, which may beformed by one or more pivot point 3620. A wing may comprise an entireportion of a base on a side of an axis. An entire portion of the basemay fold over, thereby forming the wing. In some embodiments, a centralportion 3606 of the base may intersect the axis of rotation while thewing does not. The central portion of the base may be closer to the axisof rotation than the wing. The central portion of the base may beconfigured to accept a drive mechanism 3630. The drive mechanism may bea motor, or any other mechanism that may cause the base to rotate, andmay be discussed in further detail elsewhere herein. In someembodiments, a wing may have a footprint of about 2%, 5%, 10%, 15%, 20%,25%, 30%, 35%, or 40% of the base footprint or greater.

In some embodiments, a plurality of foldover axes may be providedthrough the base. The foldover axes may be parallel to one another.Alternatively, some foldover axes may be orthogonal to one another or atany other angle relative to one another. A foldover axis may extendthrough a lower surface of the base, an upper surface of the base, orbetween the lower and upper surface of the base. In some embodiments,the foldover axis may extend through the base closer to the lowersurface of the base, or closer to the upper surface of the base. In someembodiments, a pivot point may be at or closer to a lower surface of thebase or an upper surface of the base.

One, two, three, four, five, six, or more cavities may be provided in awing. For example, a wing may be configured to accept one, two, or moresamples or sample vessels. Each wing may be capable of accepting thesame number of vessels or different numbers of vessels. The wing maycomprise a cavity configured to receive a sample vessel, wherein thesample vessel is oriented in a first orientation when the base is atrest and is configured to be oriented at a second orientation when thebase is rotating.

In some embodiments, the wing may be configured to be at angle relativeto the central portion of the base. For example, the wing may be between90 and 180 degrees of the central portion of the base. For example, thewing may be vertically oriented when the base is at rest. The wing maybe 90 degrees from the central portion of the base when verticallyoriented. The wing may be horizontally oriented when the base isrotating. The wing may be 180 degrees from the central portion of thebase when horizontally oriented. The wing may extend from the base toform a substantially uninterrupted surface when the base is rotating.For example, the wing may be extended to form a substantially continuoussurface of the bottom and/or top surface of the base when the base isrotating. The wing may be configured to fold downward relative to thecentral portion of the base.

A pivot point for a wing may include one or more pivot pin 3622. A pivotpin may extend through a portion of the wing and a portion of thecentral portion of the base. In some embodiments, the wing and centralportion of the base may have interlocking features 3624, 3626 that mayprevent the wing from sliding laterally with respect to the centralportion of the base.

A wing may have a center of gravity 3680 that is positioned lower thanthe foldover axis and/or pivot point 3620. The center of gravity of thewing may be positioned lower than the axis extending through the basewhen the base is at rest. The center of gravity of the wing may bepositioned lower than the axis extending through the base when the baseis rotating.

The wing may be formed of two or more different materials havingdifferent densities. Alternatively, the wing may be formed of a singlematerial. In one example, the wing may have a lightweight wing cap 3640and a heavy wing base 3645. In some embodiments, the wing cap may beformed of a material with a lower density than the wing base. Forexample, the wing cap may be formed of plastic while the wing base isformed of a metal, such as steel, tungsten, aluminum, copper, brass,iron, gold, silver, titanium, or any combination or alloy thereof. Aheavier wing base may assist with providing a wing center of mass belowa foldover axis and/or pivot point.

The wing cap and wing base may be connected through any mechanisms knownin the art. For example, fasteners 3650 may be provided, or adhesives,welding, interlocking features, clamps, hook and loop fasteners, or anyother mechanism may be employed. The wing may optionally include inserts3655. The inserts may be formed of a heavier material than the wing cap.The inserts may assist with providing a wing center of mass below afoldover axis and/or pivot point.

One or more cavity 3670 may be provided within the wing cap or the wingbase, or any combination thereof. In some embodiments, a cavity may beconfigured to accept a plurality of sample vessel configurations. Thecavity may have an internal surface. At least a portion of the internalsurface may contact a sample vessel. In one example, the cavity may haveone or more shelf or internal surface features that may permit a firstsample vessel having a first configuration to fit within the cavity anda second sample vessel having a second configuration to fit within thecavity. The first and second sample vessels having differentconfigurations may contact different portions of the internal surface ofthe cavity.

The centrifuge may be configured to engage with a fluid handling device.For example, the centrifuge may be configured to connect to a pipette orother fluid handling device. In some embodiments, a water-tight seal maybe formed between the centrifuge and the fluid handling device. Thecentrifuge may engage with the fluid handling device and be configuredto receive a sample dispensed from the fluid handling device. Thecentrifuge may engage with the fluid handling device and be configuredto receive a sample vessel from the fluid handling device. Thecentrifuge may engage with the fluid handling device and permit thefluid handling device to pick-up or aspirate a sample from thecentrifuge. The centrifuge may engage with the fluid handling device andpermit the fluid handling device to pick-up a sample vessel.

A sample vessel may be configured to engage with the fluid handlingdevice. For example, the sample vessel may be configured to connect to apipette or other fluid handling device. In some embodiments, awater-tight seal may be formed between the sample vessel and the fluidhandling device. The sample vessel may engage with the fluid handlingdevice and be configured to receive a sample dispensed from the fluidhandling device. The sample vessel may engage with the fluid handlingdevice and permit the fluid handling device to pick-up or aspirate asample from the sample vessel.

A sample vessel may be configured to extend out of a centrifuge wing. Insome embodiments, the centrifuge base may be configured to permit thesample vessel to extend out of the centrifuge wing when the wing isfolded over, and permit the wing to pivot between a folded and extendedstate.

FIG. 37 shows an example of a centrifuge provided in accordance withanother embodiment of the invention. The centrifuge may include a base3700 having a bottom surface 3702 and/or top surface 3704. The base maycomprise one, two or more buckets 3710 a, 3710 b.

A bucket may be configured to pivot about a bucket pivot axis extendingthrough the base. In some embodiments, the axis may form a secantthrough the base. The bucket may be configured to pivot about a point ofrotation 3720. The base may be configured to accept a drive mechanism.In one example, the drive mechanism may be a motor, such as a brushlessmotor. The drive mechanism may include a rotor 3730 and a stator 3735.The rotor may optionally be a brushless motor rotor, and the stator mayoptionally be a brushless motor stator. The drive mechanism may be anyother mechanism that may cause the base to rotate, and may be discussedin further detail elsewhere herein.

In some embodiments, a plurality of axes of rotation for the buckets maybe provided through the base. The axes may be parallel to one another.Alternatively, some axes may be orthogonal to one another or at anyother angle relative to one another. A bucket axis of rotation mayextend through a lower surface of the base, an upper surface of thebase, or between the lower and upper surface of the base. In someembodiments, the bucket axis of rotation may extend through the basecloser to the lower surface of the base, or closer to the upper surfaceof the base. In some embodiments, a point of rotation may be at orcloser to a lower surface of the base or an upper surface of the base.

One, two, three, four, or more cavities may be provided in a bucket. Forexample, a bucket may be configured to accept one, two, or more samplesor sample vessels 3740. Each bucket may be capable of accepting the samenumber of vessels or different numbers of vessels. The bucket maycomprise a cavity configured to receive a sample vessel, wherein thesample vessel is oriented in a first orientation when the base is atrest and is configured to be oriented at a second orientation when thebase is rotating.

In some embodiments, the bucket may be configured to be at anglerelative to the base. For example, the bucket may be between 0 and 90degrees of the base. For example, the bucket may be vertically orientedwhen the base is at rest. The bucket may be positioned upwards past thetop surface of the centrifuge base when the base is at rest. At least aportion of the sample vessel may extend beyond the top surface of thebase when the base is at rest. The wing may be 90 degrees from thecentral portion of the base when vertically oriented. The bucket may behorizontally oriented when the base is rotating. The bucket may be 0degrees from the base when horizontally oriented. The bucket may beretracted into the base to form a substantially uninterrupted top and/orbottom surface when the base is rotating. For example, the bucket may beretracted to form a substantially continuous surface of the bottomand/or top surface of the base when the base is rotating. The bucket maybe configured to pivot upwards relative the base. The bucket may beconfigured so that at least a portion of the bucket may pivot upwardspast the top surface of the base.

A point of rotation for a bucket may include one or more pivot pin. Apivot pin may extend through the bucket and the base. In someembodiments, the bucket may be positioned between portions of the basethat may prevent the bucket from sliding laterally with respect to thebase.

A bucket may have a center of mass 3750 that is positioned lower thanthe point of rotation 3720. The center of mass of the bucket may bepositioned lower than the point of rotation when the base is at rest.The center of mass of the bucket may be positioned lower than the pointof rotation when the base is rotating.

The bucket may be formed of two or more different materials havingdifferent densities. Alternatively, the bucket may be formed of a singlematerial. In one example, the bucket may have a main body 3715 and an ininsert 3717. In some embodiments, the main body may be formed of amaterial with a lower density than the insert. For example, the mainbody may be formed of plastic while the insert is formed of a metal,such as tungsten, steel, aluminum, copper, brass, iron, gold, silver,titanium, or any combination or alloy thereof. A heavier insert mayassist with providing a bucket center of mass below a point of rotation.The bucket materials may include a higher density material and a lowerdensity material, wherein the higher density material is positionedlower than the point of rotation. The center of mass of the bucket maybe located such that the bucket naturally swings with an open endupwards, and heavier end downwards when the centrifuge is at rest. Thecenter of mass of the bucket may be located so that the bucket naturallyretracts when the centrifuge is rotated at a certain speed. The bucketmay retract when the speed is at a predetermined speed, which mayinclude any speed, or any speed mentioned elsewhere.

One or more cavity may be provided within the bucket. In someembodiments, a cavity may be configured to accept a plurality of samplevessel configurations. The cavity may have an internal surface. At leasta portion of the internal surface may contact a sample vessel. In oneexample, the cavity may have one or more shelf or internal surfacefeatures that may permit a first sample vessel having a firstconfiguration to fit within the cavity and a second sample vessel havinga second configuration to fit within the cavity. The first and secondsample vessels having different configurations may contact differentportions of the internal surface of the cavity.

As previously described, the centrifuge may be configured to engage witha fluid handling device. For example, the centrifuge may be configuredto connect to a pipette or other fluid handling device. The centrifugemay be configured to accept a sample dispensed by the fluid handlingdevice or to provide a sample to be aspirated by the fluid handlingdevice. A centrifuge may be configured to accept or provide a samplevessel.

A sample vessel may be configured to engage with the fluid handlingdevice, as previously mentioned. For example, the sample vessel may beconfigured to connect to a pipette or other fluid handling device.

A sample vessel may be configured to extend out of a bucket. In someembodiments, the centrifuge base may be configured to permit the samplevessel to extend out of the bucket when the bucket is provided in aretracted state, and permit the bucket to pivot between a retracted andprotruding state. The sample vessel extending out of the top surface ofthe centrifuge may permit easier sample or sample vessel transfer toand/or from the centrifuge. In some embodiments, the buckets may beconfigured to retract into the rotor, creating a compact assembly andreducing drag during operation, with additional benefits such as reducednoise and heat generation, and lower power requirements.

In some embodiments, the centrifuge base may include one or morechannels, or other similar structures, such as grooves, conduits, orpassageways. Any description of channels may also apply to any of thesimilar structures. The channels may contain one or more ball bearing.The ball bearings may slide through the channels. The channels may beopen, closed, or partially open. The channels may be configured toprevent the ball bearings from falling out of the channel.

In some embodiments, ball bearings may be placed within the rotor in asealed/closed track. This configuration is useful for dynamicallybalancing the centrifuge rotor, especially when centrifuging samples ofdifferent volumes at the same time. In some embodiments, the ballbearings may be external to the motor, making the overall system morerobust and compact.

The channels may encircle the centrifuge base. In some embodiments, thechannel may encircle the base along the perimeter of the centrifugebase. In some embodiments, the channel may be at or closer to an uppersurface of the centrifuge base, or the lower surface of the centrifugebase. In some instances, the channel may be equidistant to the upper andlower surface of the centrifuge base. The ball bearings may slide alongthe perimeter of the centrifuge base. In some embodiments, the channelmay encircle the base at some distance away from the axis rotation. Thechannel may form a circle with the axis of rotation at the substantialcenter of the circle.

FIG. 38 shows an additional example of a centrifuge provided inaccordance with another embodiment of the invention. The centrifuge mayinclude a base 3800 having a bottom surface 3802 and/or top surface3804. The base may comprise one, two or more buckets 3810 a, 3810 b. Abucket may be connected to a module frame 3820 which may be connected tothe base. Alternatively, the bucket may directly connect to the base.The bucket may also be attached to a weight 3830.

A module frame may be connected to a base. The module frame may beconnected to the base at a boundary that may form a continuous orsubstantially continuous surface with the base. A portion of the top,bottom and/or side surface of the base may form a continuous orsubstantially continuous surface with the module frame.

A bucket may be configured to pivot about a bucket pivot axis extendingthrough the base and/or module frame. In some embodiments, the axis mayform a secant through the base. The bucket may be configured to pivotabout a bucket pivot 3840. The base may be configured to accept a drivemechanism. In one example, the drive mechanism may be a motor, such as abrushless motor. The drive mechanism may include a rotor 3850 and astator 3855. In some embodiments, the rotor may be a brushless motorrotor, and the stator may be a brushless motor stator. The drivemechanism may be any other mechanism that may cause the base to rotate,and may be discussed in further detail elsewhere herein.

In some embodiments, a plurality of axes of rotation for the buckets maybe provided through the base. The axes may be parallel to one another.Alternatively, some axes may be orthogonal to one another or at anyother angle relative to one another. A bucket axis of rotation mayextend through a lower surface of the base, an upper surface of thebase, or between the lower and upper surface of the base. In someembodiments, the bucket axis of rotation may extend through the basecloser to the lower surface of the base, or closer to the upper surfaceof the base. In some embodiments, a bucket pivot may be at or closer toa lower surface of the base or an upper surface of the base. A bucketpivot may be at or closer to a lower surface of the module frame or anupper surface of the module frame.

One, two, three, four, or more cavities may be provided in a bucket. Forexample, a bucket may be configured to accept one, two, or more samplesor sample vessels. Each bucket may be capable of accepting the samenumber of vessels or different numbers of vessels. The bucket maycomprise a cavity configured to receive a sample vessel, wherein thesample vessel is oriented in a first orientation when the base is atrest and is configured to be oriented at a second orientation when thebase is rotating.

In some embodiments, the bucket may be configured to be at an anglerelative to the base. For example, the bucket may be between 0 and 90degrees of the base. For example, the bucket may be vertically orientedwhen the base is at rest. The bucket may be positioned upwards past thetop surface of the centrifuge base when the base is at rest. At least aportion of the sample vessel may extend beyond the top surface of thebase when the base is at rest. The wing may be 90 degrees from thecentral portion of the base when vertically oriented. The bucket may behorizontally oriented when the base is rotating. The bucket may be 0degrees from the base when horizontally oriented. The bucket may beretracted into the base and/or frame module to form a substantiallyuninterrupted top and/or bottom surface when the base is rotating. Forexample, the bucket may be retracted to form a substantially continuoussurface with the bottom and/or top surface of the base and/or framemodule when the base is rotating. The bucket may be configured to pivotupwards relative the base and/or frame module. The bucket may beconfigured so that at least a portion of the bucket may pivot upwardspast the top surface of the base and/or frame module.

The bucket may be locked in multiple positions to enable drop-off andpickup of centrifuge tubes, as well as aspiration and dispensing ofliquid into and out of a centrifuge vessel when in the centrifugebucket. One means to accomplish this is one or more motors that drivewheels that make contact with the centrifuge rotor to finely positionand/or lock the rotor. Another approach may be to use a CAM shape formedon the rotor, without additional motors or wheels. An appendage from thepipette, such as a centrifuge tip attached to a pipette nozzle, may bepressed down onto the CAM shape on the rotor. This force on the CAMsurface may induce the rotor to rotate to the desired locking position.The continued application of this force may enable the rotor to berigidly held in the desired position. Multiple such CAM shapes may beadded to the rotor to enable multiple locking positions. While the rotoris held by one pipette nozzle/tip, another pipette nozzle/tip mayinterface with the centrifuge buckets to drop off or pick up centrifugevessels or perform other functions, such as aspirating or dispensingfrom the centrifuge vessels in the centrifuge bucket.

A bucket pivot may include one or more pivot pin. A pivot pin may extendthrough the bucket and the base and/or frame module. In someembodiments, the bucket may be positioned between portions of the baseand/or frame module that may prevent the bucket from sliding laterallywith respect to the base.

The bucket may be attached to a weight. The weight may be configured tomove when the base starts rotating, thereby causing the bucket to pivot.The weight may be caused to move by a centrifugal force exerted on theweight when the base starts rotating. The weight may be configured tomove away from an axis of rotation when the base starts rotating at athreshold speed. In some embodiments, the weight may move in a lineardirection or path. Alternatively, the weight may move along a curvedpath or any other path. The bucket may be attached to a weight at aweight pivot point 3860. One or more pivot pin or protrusion may be usedthat may allow the bucket to rotate with respect to the weight. In someembodiments, the weight may move along a horizontal linear path, therebycausing the bucket to pivot upward or downward. The weight may move in alinear direction orthogonal to the axis of rotation of the centrifuge.

The weight may be located between portions of a module frame and/or abase. The module frame and/or base may be configured to prevent theweight from sliding out of the base. The module and/or base may restrictthe path of the weight. The path of the weight may be restricted to alinear direction. One or more guide pins 3870 may be provided that mayrestrict the path of the weight. In some embodiments, the guide pins maypass through the frame module and/or base and the weight.

A biasing force may be provided to the weight. The biasing force may beprovided by a spring 3880, elastic, pneumatic mechanism, hydraulicmechanism, or any other mechanism. The biasing force may keep the weightat a first position when the base is at rest, while the centrifugalforce from the rotation of the centrifuge may cause the weight to moveto a second position when the centrifuge is rotating at a thresholdspeed. When the centrifuge goes back to rest or the speed falls below apredetermined rotation speed, the weight may return to the firstposition. The bucket may have a first orientation when the weight is atthe first position, and the bucket may have a second orientation whenthe weight is at the second position. For example, the bucket may have avertical orientation when the weight is in the first position and thebucket may have a horizontal orientation when the weight is in thesecond position. The first position of the weight may be closer to theaxis of rotation than the second position of the weight.

One or more cavity may be provided within the bucket. In someembodiments, a cavity may be configured to accept a plurality of samplevessel configurations. The cavity may have an internal surface. At leasta portion of the internal surface may contact a sample vessel. In oneexample, the cavity may have one or more shelf or internal surfacefeatures that may permit a first sample vessel having a firstconfiguration to fit within the cavity and a second sample vessel havinga second configuration to fit within the cavity. The first and secondsample vessels having different configurations may contact differentportions of the internal surface of the cavity.

As previously described, the centrifuge may be configured to engage witha fluid handling device. For example, the centrifuge may be configuredto connect to a pipette or other fluid handling device. The centrifugemay be configured to accept a sample dispensed by the fluid handlingdevice or to provide a sample to be aspirated by the fluid handlingdevice. A centrifuge may be configured to accept or provide a samplevessel.

A sample vessel may be configured to engage with the fluid handlingdevice, as previously mentioned. For example, the sample vessel may beconfigured to connect to a pipette or other fluid handling device.

A sample vessel may be configured to extend out of a bucket. In someembodiments, the centrifuge base and/or module frame may be configuredto permit the sample vessel to extend out of the bucket when the bucketis provided in a retracted state, and permit the bucket to pivot betweena retracted and protruding state. The sample vessel extending out of thetop surface of the centrifuge may permit easier sample or sample vesseltransfer to and/or from the centrifuge.

In some embodiments, the centrifuge base may include one or morechannels, or other similar structures, such as grooves, conduits, orpassageways. Any description of channels may also apply to any of thesimilar structures. The channels may contain one or more ball bearing.The ball bearings may slides through the channels. The channels may beopen, closed, or partially open. The channels may be configured toprevent the ball bearings from falling out of the channel.

The channels may encircle the centrifuge base. In some embodiments, thechannel may encircle the base along the perimeter of the centrifugebase. In some embodiments, the channel may be at or closer to an uppersurface of the centrifuge base, or the lower surface of the centrifugebase. In some instances, the channel may be equidistant to the upper andlower surface of the centrifuge base. The ball bearings may slide alongthe perimeter of the centrifuge base. In some embodiments, the channelmay encircle the base at some distance away from the axis rotation. Thechannel may form a circle with the axis of rotation at the substantialcenter of the circle.

Other examples of centrifuge configurations known in the art, includingvarious swinging bucket configurations, may be used. See, e.g., U.S.Pat. No. 7,422,554 which is hereby incorporated by reference in itsentirety. For examples, buckets may swing down, rather than swinging up.Buckets may swing to protrude to the side rather than up or down.

The centrifuge may be enclosed within a housing or casing. In someembodiments, the centrifuge may be completely enclosed within thehousing. Alternatively, the centrifuge may have one or more opensections. The housing may include a movable portion that may allow afluid handling or other automated device to access the centrifuge. Thefluid handling and/or other automated device may provide a sample,access a sample, provide a sample vessel, or access a sample vessel in acentrifuge. Such access may be granted to the top, side, and/or bottomof the centrifuge.

A sample may be dispensed and/or picked up from the cavity. The samplemay be dispensed and/or picked up using a fluid handling system. Thefluid handling system may be the pipette described elsewhere herein, orany other fluid handling system known in the art. The sample may bedispensed and/or picked up using a tip, having any of the configurationsdescribed elsewhere herein. The dispensing and/or aspiration of a samplemay be automated.

In some embodiments, a sample vessel may be provided to or removed froma centrifuge. The sample vessel may be inserted or removed from thecentrifuge using a device in an automated process. The sample vessel mayextend from the surface of the centrifuge, which may simplify automatedpick up and/or retrieval. A sample may already be provided within thesample vessel. Alternatively, a sample may be dispensed and/or picked upfrom the samples vessel. The sample may be dispensed and/or picked upfrom the sample vessel using the fluid handling system.

In some embodiments, a tip from the fluid handling system may beinserted at least partially into the sample vessel and/or cavity. Thetip may be insertable and removable from the sample vessel and/orcavity. In some embodiments the sample vessel and the tip may be thecentrifugation vessel and centrifugation tip as previously described, orhave any other vessel or tip configuration. In some embodiments, acuvette, such as described in FIGS. 70A and 70B can be placed in thecentrifuge rotor. This configuration may offer certain advantages overtraditional tips and/or vessels. In some embodiments, the cuvettes maybe patterned with one or more channels with specialized geometries suchthat products of the centrifugation process are automatically separatedinto separate compartments. One such embodiment might be a cuvette witha tapered channel ending in a compartment separated by a narrow opening.The supernatant (e.g. plasma from blood) can be forced into thecompartment by centrifugal forces, while the red blood cells remain inthe main channel. The cuvette may be more complicated with severalchannels and/or compartments. The channels may be either isolated orconnected.

In some embodiments, one or more cameras may be placed in the centrifugerotor such that it can image the contents of the centrifuge vessel whilethe rotor is spinning. The camera images may be analyzed and/orcommunicated in real time, such as by using a wireless communicationmethod. This method may be used to track the rate of sedimentation/cellpacking, such as for the ESR (erythrocyte sedimentation rate) assay,where the speed of RBC (red blood cell) settling is measured. In someembodiments, one or more cameras may be positioned outside the rotorthat can image the contents of the centrifuge vessel while the rotor isspinning. This may be achieved by using a strobed light source that istimed with the camera and spinning rotor. Real-time imaging of thecontents of a centrifuge vessel while the rotor is spinning may allowone to stop spinning the rotor after the centrifugation process hascompleted, saving time and possibly preventing over-packing and/orover-separation of the contents.

Referring now to FIG. 94, one embodiment of a centrifuge with a sampleimaging system will now be described. FIG. 94 shows that, in someembodiments, the imaging device 3750 such as but not limited to acamera, a CCD sensor, or the like may be used with a centrifuge rotor3800. In this example, the imaging device 3750 is stationary while thecentrifuge rotor 3800 is spinning Imaging may be achieved by using astrobed light source that is timed with the camera and spinning rotor.Optionally, high speed image capture can also be used to acquire imageswithout the use of a strobe.

FIG. 95 shows one embodiment of the imaging device 3750 that can bemounted in a stationary position to view the centrifuge vessel while itis spinning in the centrifuge. FIG. 95 shows that in addition to theimaging device 3750, illumination source(s) 3752 and 3754 may also beused to assist in image capture. The mounting device 3756 is configuredto position the imaging device 3750 to have a field of view and focusthat enables a clear view of the centrifuge vessel and content therein.

Referring now to FIGS. 96 to 98, yet another embodiment of a centrifugewith a sample imaging system will now be described. FIG. 96 shows that,in some embodiments, the imaging device 3770 such as but not limited toa camera, a CCD sensor, or the like may be mounted inside or in the samerotation frame of reference as the centrifuge rotor 3800. FIG. 97 showsa cross-sectional view showing that the imaging device 3770 ispositioned to view into the sample in the centrifuge vessel 3772 throughan opening 3774 (shown in FIG. 98). Because the imaging system is in thecentrifuge rotor 3800, the imaging system can continuously image thecentrifuge vessel 3772 and the sample therein without the use of astrobe illumination system. Optionally, the centrifuge rotor 3800 can beappropriately balanced to account for the additional weight of theimaging device 3770 in the rotor.

Thermal Control Unit

In accordance with some embodiments of the invention, a system mayinclude one or more thermal control unit. A device may include one ormore thermal control unit therein. For example, one or more thermalcontrol unit may be provided within a device housing. A module may haveone or more thermal control unit. One, two, or more modules of a devicemay have a thermal control unit therein. The thermal control unit may besupported by a module support structure, or may be contained within amodule housing. A thermal control unit may be provided at a device level(e.g., overall across all modules within a device), rack level (e.g.,overall across all modules within a rack), module level (e.g., within amodule), and/or component level (e.g., within one or more components ofa module).

A thermal control unit may be configured to heat and/or cool a sample orother fluid or module temperature or temperature of the entire device.Any discussion of controlling the temperature of a sample may also referto any other fluid herein, including but not limited to reagents,diluents, dyes, or wash fluid. In some embodiments, separate thermalcontrol unit components may be provided to heat and cool the sample.Alternatively, the same thermal control unit components may both heatand cool the sample.

The thermal control unit may be used to vary and/or maintain thetemperature of a sample to keep the sample at a desire temperature orwithin a desired temperature range. In some embodiments, the thermalcontrol unit may be capable of maintaining the sample within 1 degree C.of a target temperature. In other embodiments, the thermal control unitmay be capable of maintaining the sample within 5 degrees C., 4 degreesC., 3 degrees C., 2 degrees C., 1.5 degrees C., 0.75 degrees C., 0.5degrees C., 0.3 degrees C., 0.2 degrees C., 0.1 degrees C., 0.05 degreesC., or 0.01 degrees C. of the target temperature. A desired targettemperature may be programmed. The desired target temperature may varyor may be maintained over time. A target temperature profile may accountfor variations in desired target temperature over time. The targettemperature profile may be provided dynamically from an external device,such as a server, may be provided from on-board the device, or may beentered by an operator of the device.

The thermal control unit may be able to account for temperaturesexternal to the device. For example, one or more temperature sensor maydetermine the environmental temperature external to the device. Thethermal control unit may operate to reach a target temperature,compensating for different external temperatures.

The target temperature may remain the same or may vary over time. Insome embodiments, the target temperature may vary in a cyclic manner. Insome embodiments, the target temperature may vary for a while and thenremain the same. In some embodiments, the target temperature may followa profile as known in the art for nucleic acid amplification. Thethermal control unit may control the sample temperature to follow theprofile known for nucleic acid amplification. In some embodiments, thetemperature may be in the range of about 30-40 degrees Celsius. In someinstances, the range of temperature is about 0-100 degrees Celsius. Forexample, for nucleic acid assays, temperatures up to 100 degrees Celsiuscan be achieved. In an embodiment, the temperature range is about 15-50degrees Celsius. In some embodiments, the temperature may be used toincubate one or more sample.

The thermal control unit may be capable of varying the temperature ofone or more sample quickly. For example, the thermal control unit mayramp the sample temperature up or down at a rate of more than and/orequal to 1 C/min, 5 C/min, 10 C/min, 15 C/min, 30 C/min, 45 C/min, 1C/sec, 2 C/sec, 3 C/sec, 4 C/sec, 5 C/sec, 7 C/sec, or 10 C/sec.

A thermal control unit of the system can comprise a thermoelectricdevice. In some embodiments, the thermal control unit can be a heater. Aheater may provide active heating. In some embodiments, voltage and/orcurrent provided to the heater may be varied or maintained to provide adesired amount of heat. A thermal control unit may be a resistiveheater. The heater may be a thermal block. In one embodiment, a thermalblock is used in a nucleic acid assay station to regulate thetemperature of reactions.

A thermal block may have one or many openings to enable incorporation ofdetectors and/or light sources. Thermal blocks may have openings forimaging of contents. Openings in thermal blocks can be filled and/orcovered to improve thermal properties of the block.

The heater may or may not have components that provide active cooling.In some embodiments, the heater may be in thermal communication with aheat sink. The heat sink may be passively cooled, and may permit heat todissipate to the surrounding environment. Is some embodiments, the heatsink or the heater may be actively cooled, such as with forced fluidflow. The heat sink may or may not contain one or more surface featuresuch as fins, ridges, bumps, protrusions, grooves, channels, holes,plates, or any other feature that may increase the surface area of theheat sink. In some embodiments, one or more fan or pump may be used toprovide forced fluid cooling.

In some embodiments, the thermal control unit can be a Peltier device ormay incorporate a Peltier device.

The thermal control unit may optionally incorporate fluid flow toprovide temperature control. For example, one or more heated fluid orcooled fluid may be provided to the thermal control unit. In someembodiments, heated and/or cooled fluid may be contained within thethermal control unit or may flow through the thermal control unit. Airtemperature control can be enhanced by the use of heat pipes to rapidlyraise temperature to a desired level. By using forced convection, heattransfer can be made faster. Forced convective heat transfer could alsobe used to thermocycle certain regions by alternately blowing hot andcold air. Reactions requiring specific temperatures and temperaturecycling can be done on a tip and/or vessel, where heating and cooling ofthe tip is finely controlled, such as by an IR heater.

In some embodiments, a thermal control unit may use conduction,convection and/or radiation to provide heat to, or remove heat from asample. In some embodiments, the thermal control unit may be in directphysical contact with a sample or sample holder. The thermal controlunit may be in direct physical contact with a vessel, tip, microcard, orhousing for a vessel, tip, or microcard. The thermal control unit maycontact a conductive material that may be in direct physical contactwith a sample or sample holder. For example, the thermal control unitmay contact a conductive material that may be in direct physical contactwith a vessel, tip, microcard, or a housing to support a vessel, tip, ormicrocard. In some embodiments, the thermal control unit may be formedof or include a material of high thermal conductivity. For example, thethermal control unit may include a metal such as copper, aluminum,silver, gold, steel, brass, iron, titanium, nickel or any combination oralloy thereof. For example, the thermal control unit can include a metalblock. In some embodiments, the thermal control unit may include aplastic or ceramic material.

One or more samples may be brought to and/or removed from the thermalcontrol unit. In some embodiments, the samples may be brought to and/orremoved from the thermal control unit using a fluid handling system. Thesamples may be brought to and/or removed from the thermal control unitusing any other automated process. The samples may be transported to andfrom the thermal control unit without requiring human intervention. Insome embodiments, the samples may be manually transferred to or from thethermal control unit.

The thermal control unit may be configured to be in thermalcommunication with a sample of a small volume. For example, the thermalcontrol unit may be configured to be thermal communication with a samplewith a volume as described elsewhere herein.

The thermal control unit may be in thermal communication with aplurality of samples. In some instances, the thermal control unit maykeep each of the same samples at the same temperature relative to oneanother. In some instances, a thermal control unit may be thermallyconnected to a heat spreader which may evenly provide heat to theplurality of samples.

In other embodiments, the thermal control unit may provide differentamounts of heat to the plurality of samples. For example, a first samplemay be kept at a first target temperature, and a second sample may bekept at a second target temperature. The thermal control unit may form atemperature gradient. In some instances, separate thermal control unitsmay keep different samples at different temperatures, or operating alongseparate target temperature profiles. A plurality of thermal controlunits may be independently operable.

One or more sensor may be provided at or near the thermal control unit.One or more sensor may be provided at or near a sample in thermalcommunication with the thermal control unit. In some embodiments, thesensor may be a temperature sensor. Any temperature sensor known in theart may be used including, but not limited to thermometers,thermocouples, or IR sensors. A sensor may provide one or more signal toa controller. Based on the signal, the controller may send a signal tothe thermal control unit to modify (e.g., increase or decrease) ormodify the temperature of the sample. In some embodiments, thecontroller may directly control the thermal control unit to modify ormaintain the sample temperature. The controller may be separate from thethermal control unit or may be a part of the thermal control unit.

In some embodiments, the sensors may provide a signal to a controller ona periodic basis. In some embodiments, the sensors may provide real-timefeedback to the controller. The controller may adjust the thermalcontrol unit on a periodic basis or in real-time in response to thefeedback.

As previously mentioned, the thermal control unit may be used fornucleic acid amplification (e.g., isothermal and non-isothermal nucleicacid amplification, such as PCR), incubation, evaporation control,condensation control, achieving a desired viscosity, separation, or anyother use known in the art.

Nucleic Acid Assay Station

In some embodiments, a system, device, or module disclosed herein maycontain a nucleic acid assay station. A nucleic acid assay station maycontain one or more hardware components for facilitating the performanceof nucleic acid assays (e.g. a thermal control unit). A nucleic acidassay station may also contain one or more detection units or sensorsfor monitoring or measuring non-nucleic acid assays (e.g. generalchemistry assays, immunoassays, etc.). A nucleic acid assay station maybe incorporated with or may be separate from a cartridge or generalassay station of a module or device. A nucleic acid assay station mayalso be referred herein to as a “nucleic acid amplification module.”

FIG. 101 shows an example of a nucleic acid assay station 10201. Anucleic acid assay station 10201 may contain a thermal block 10202. Thethermal block 10202 may be shaped to receive or support one or morevessels 10203 (including assay units, tips, and any nucleic acidvessel/tip disclosed elsewhere herein), such as by having wells. Thethermal block may have any of the features of a thermal control unitdescribed elsewhere herein. For example, the thermal block may maintaina selected temperature or range or cycle of temperatures in order toperform or support a nucleic acid assay (e.g. to thermocycle for a PCRassay or to maintain a selected constant temperature for an isothermalassay). In some embodiments, the thermal block may be in thermal contactwith a heater or thermal control unit, such that the thermal blockitself does not contain components for regulating heat. Instead, thetemperature of the thermal block may be regulated by the temperature ofthe heater or thermal control unit in thermal contact with the heatingblock.

A nucleic acid assay station may be configured to receive 1, 2, 3, 4, 5,6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 35, 40, 45, 50,60, 70, 80, 90, 100, 125, 150, 175, 200, 225, 250, 275, 300, 400, 500,or more vessels. In some embodiments, a thermal block may contain 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 35, 40,45, 50, 60, 70, 80, 90, 100, 125, 150, 175, 200, 225, 250, 275, 300,400, 500, or more wells. A nucleic acid assay station may be positionedin a device or module such that it may be accessed by a sample handlingsystem of the device or module. For example, a sample handling system ofa device or module may be configured to transport vessels to or from anucleic acid assay station.

In some embodiments, a nucleic acid assay station 10201 may contain amovable portion 10204. The moveable portion may be configured formovement along a guide structure of the station, such as a track 10205or guide rod. The moveable portion may have two or more positions,including an open position and a closed position. When the movableportion 10204 is in an open position, the wells of a thermal block 10202may be accessible, so that vessels may be placed in or removed from thethermal block (e.g. by a sample handling system). In contrast, when themovable portion 10204 is in a closed position, it may obstruct one ormore wells of the thermal block 10202, such that vessels cannot beplaced in or removed from the thermal block.

In some embodiments, a nucleic acid assay station may contain one ormore light sources. In some embodiments, a nucleic acid assay stationmay contain the same number of light sources as number of vessels as thestation is configured to receive (e.g. if the station is configured toreceive 10 vessels, it contains 10 light sources). The light source maybe any light source disclosed elsewhere herein, including, for example alaser or a light-emitting diode. The light source(s) may be configuredsuch that it is in a fixed position relative to a thermal block orvessel wells. A light source may be in-line with a well of the thermalblock, or it may be to the side (e.g. at a 90 degree angle).Alternatively, the light source(s) may be movable relative to thethermal block or other components of the nucleic acid assay station. Thelight source(s) may be supported by a moveable portion of the station.In some embodiments, when the movable portion is in a closed position,light sources(s) supported by the movable portion are positioned suchthat light from the light source(s) is directed to the wells of athermal block or the vessels therein. In some embodiments, one or morecomponents of the nucleic acid assay station may be moveable relative tothe light source.

In some embodiments, a nucleic acid assay station may contain one ormore optical sensors. In some embodiments, a nucleic acid assay stationmay contain the same number of optical sensors as number of vessels asthe station is configured to receive (e.g. if the station is configuredto receive 10 vessels, it contains 10 optical sensors). The opticalsensor may be any sensor for detecting light signals disclosed elsewhereherein, including, for example a PMT, photodiode, or CCD sensor. Theoptical sensor may be configured such that it is in a fixed positionrelative to a thermal block or vessel wells. An optical sensor may bein-line with a well of the thermal block, or it may be to the side (e.g.at a 90 degree angle). Alternatively, the optical sensors(s) may bemovable relative to the thermal block or other components of the nucleicacid assay station. The optical sensors (s) may be supported by amoveable portion of the station. In some embodiments, when the movableportion is in a closed position, optical sensors (s) supported by themovable portion are positioned such that light generated from or passingthrough the wells of a thermal block or the vessels therein may reachthe optical sensor. In some embodiments, one or more components of thenucleic acid assay station may be moveable relative to the opticalsensor.

A nucleic acid assay station may contain both a light source and anoptical sensor. Stations containing both a light source and an opticalsensor may have capabilities similar to a spectrophotometer. In someembodiments, a nucleic acid assay station containing both a light sourceand optical sensor may be configured to perform a measurement involvingassessing an optical property of a sample which is typically performedin a dedicated spectrophotometer—for example, measurement of: color,absorbance, transmittance, fluorescence, light-scattering properties, orturbidity of a sample. In some embodiments, a nucleic acid assay stationcontaining both a light source and optical sensor can perform ameasurement of a sample that only uses the optical sensor—e.g.measurement of the luminescence of a sample. In such situations, thelight source of the station may be turned off or blocked while theoptical sensor detects light emitted from the sample. Assay types thatmay be measured include, for example, nucleic acid assays, immunoassays,and general chemistry assays.

In some embodiments, a nucleic acid assay station may contain an opticalsensor and optionally, a light source for each well of the heating blockor station. Inclusion of an optical sensor for each well may permit thesimultaneous measurement of multiple different assays in the nucleicacid assay station at the same time.

In some embodiments, nucleic acid assay station may contain an opticalsensor at a fixed position in or adjacent to the thermal block. Theoptical sensor may be in-line with the well of a thermal block, or tothe side of the well of the thermal block. There may be an opening or achannel in the wall of the well of thermal block creating an opticalpath between the interior of the well and the optical sensor. Thenucleic acid assay station may also contain a light source. The lightsource may be attached to a movable portion of the assay station,configured such that in one or more positions of the movable portion,the light from the light source is directed into the well of the thermalblock. In situations where the light source and the optical sensor areboth in-line with the well of the thermal block (due to the light sourceand optical sensor having fixed or movable positions), various types ofspectrophotometric readings of the sample may be obtained—e.g.absorbance, transmittance, or fluorescence. In situations where theoptical sensor is at an angle to the light source and the well of thethermal block, spectrophotometric readings of the sample that may beobtained include, for instance, light scattering, fluorescence, andturbidity.

To perform fluorescence assays in a nucleic acid assay station, a lightsource having a narrow emission wavelength profile may be used (e.g. alight emitting diode). In addition or alternatively, an excitationfilter may be placed between the light source and the sample, such thatlight of only a selected wavelength(s) reaches the sample. Furthermore,an emissions filter may be placed between the sample and the opticaldetector, such that only light of a selected wavelength (typically thatwhich is emitted by the fluorescent compound) reaches the opticaldetector.

In some embodiments, a nucleic acid assay (e.g. a nucleic acidamplification assay) may be performed or detected in a nucleic acidassay station. Given the various optical configurations of nucleic acidassay stations, the stations can be configured to measure nucleic acidamplification assays which result in multiple different types of opticalchanges in the reaction, such as fluorescence or turbidity. In addition,in some embodiments, any type of assay resulting in a change in opticalproperties of the sample may be measured in a nucleic acid assaystation. For example, a non-nucleic acid assay resulting in a change ofturbidity of a sample may be measured in a nucleic acid assay station,by measuring, for example, the absorbance of the sample or the lightscattered by the sample. In some embodiments, a nucleic acid assaystation may have certain wells of the thermal block configured formeasurement of fluorescence of samples (e.g. they may contain filters orlight sources of particular wavelengths), and certain wells of thethermal block configured for measurement of turbidity of samples (e.g.they may have optical sensors at an angle to the light source and wellor they may lack filters). In some embodiments, a nucleic acid assaystation may have one or more wells that are configured for detectingnucleic acid assays, and one or more wells that are configured fordetecting non-nucleic acid assays.

In some embodiments, assay units or other reaction vessels describedelsewhere herein may be transported to or situated in a nucleic acidassay station described herein for measurement of the reaction in thevessel. Accordingly, in addition to supporting nucleic acid assays, anucleic acid assay station may function as a detection unit for a widerange of assays (e.g. immunoassays and general chemistry assays). Thismay facilitate performing and detecting multiple different assayssimultaneously in a module or device provided herein.

Cytometer

In accordance with some embodiments of the invention, a system mayinclude one or more cytometer. A device may include one or morecytometer therein. For example, one or more cytometer may be providedwithin a device housing. A module may have one or more cytometer. One,two, or more modules of a device may have a cytometer therein. Thecytometer may be supported by a module support structure, or may becontained within a module housing. Alternatively, the cytometer may beprovided external to the module. In some instances, a cytometer may beprovided within a device and may be shared by multiple modules. Thecytometer may have any configuration known or later developed in theart.

In some embodiments, the cytometer may have a small volume. For example,the cytometer may have a volume of less than or equal to about 0.1 mm³,0.5 mm³, 1 mm³, 3 mm³, 5 mm³, 7 mm³, 10 mm³, 15 mm³, 20 mm³, 25 mm³, 30mm³, 40 mm³, 50 mm³, 60 mm³, 70 mm³, 80 mm³, 90 mm³, 100 mm³, 125 mm,150 mm³, 200 mm³, 250 mm³, 300 mm³, 500 mm³, 750 mm³, or 1 m³.

The cytometer may have a footprint of about less than or equal to 0.1mm², 0.5 mm², 1 mm², 3 mm², 5 mm², 7 mm², 10 mm², 15 mm², 20 mm², 25mm², 30 mm², 40 mm², 50 mm², 60 mm², 70 mm², 80 mm², 90 mm², 100 mm²,125 mm², 150 mm², 200 mm², 250 mm², 300 mm², 500 mm², 750 mm², or 1 m².The cytometer may have one or more dimension (e.g., width, length,height) of less than or equal to 0.05 mm, 0.1 mm, 0.5 mm, 0.7 mm, 1 mm,2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 7 mm, 8 mm, 9 mm, 10 mm, 11 mm, 12 mm, 13mm, 15 mm, 17 mm, 20 mm, 25 mm, 30 mm, 40 mm, 50 mm, 60 mm, 70 mm, 80mm, 100 mm, 150 mm, 200 mm, 300 mm, 500 mm, or 750 mm.

The cytometer may accept a small volume of sample or other fluid. Forexample, the cytometer may accept a volume of sample of about 500 μL orless, 250 μL or less, 200 μL or less, 175 μL or less, 150 μL or less,100 μL or less, 80 μL or less, 70 μL or less, 60 μL or less, 50 μL orless, 30 μL or less, 20 μL or less, 15 μL or less, 10 μL or less, 8 μLor less, 5 μL or less, 1 μL or less, 500 nL or less, 300 nL or less, 100nL or less, 50 nL or less, 10 nL or less, 1 nL or less, 500 pL or less,250 pL or less, 100 pL or less, 50 pL or less, 10 pL or less, 5 pL orless, or 1 pL or less.

The cytometer may utilize one or more illumination techniques, includingbut not limited to bright field, dark field, forward illumination,oblique illumination, back illumination, phase contrast and differentialinterference contrast microscopy. Focusing may be achieved using any ofthe illumination sources, including but not limited to dark fieldimaging. Dark field imaging may be performed with a various illuminationsources of different wavelength bands. Dark field imaging may beperformed with a light guide outside the objective. Images produced bythe imaging system may be monochromatic and/or color. The imaging systemmay be configured to be optics free, reducing cost and size.

The cytometer (as well as other modules) can be configured toincorporate image processing algorithms to extract quantitativeinformation from images of cells and other elements in the sample toenable computation of reportables. Where employed, the image processingand analysis may include but are not limited to: a) image acquisition,compression/decompression and quality improvement, b) imagesegmentation, c) image stitching, and d) extraction of quantitativeinformation.

Detection Unit

In accordance with some embodiments of the invention, a system mayinclude one or more detection unit. In some embodiments, a detectionstation provided herein may contain a detection unit. A device mayinclude one or more detection unit therein. For example, one or moredetection unit may be provided within a device housing. A module mayhave one or more detection unit. One, two, or more modules of a devicemay have a detection unit therein. The detection unit may be supportedby a module support structure, or may be contained within a modulehousing. Alternatively, the detection unit may be provided external tothe module.

The detection unit may be used to detect a signal produced by at leastone assay on the device. The detection unit may be used to detect asignal produced at one or more sample preparation station in a device.The detection unit may be capable of detecting a signal produced at anystage in a sample preparation or assay of the device.

In some embodiments, a plurality of detection units may be provided. Theplurality of detection units may operate simultaneously and/or insequence. The plurality of detection units may include the same types ofdetection units and/or different types of detection units. The pluralityof detection units may operate on a synchronized schedule orindependently of one another.

In some embodiments, systems, devices, or modules provided herein mayhave multiple types of detection units, which may be in one or moredetection stations. For example, a system, device or module providedherein may contain one or more, two or more, three or more, or all fourof: i) a dedicated spectrophotometer (for example, a spectrophotometeras described in FIG. 74); ii) a light sensor which is not speciallyconfigured to operate with a light source (for example, a PMT orphotodiode which is not part of a spectrophotometer); iii) a camera (forexample, containing a CCD or CMOS sensor); and iv) a nucleic acid assaystation containing or operatively coupled to a light source and a lightsensor, such that it may function as a spectrophotometer. In someembodiments, a system, device, or module provided herein may furthercontain a cytometry station containing an imaging device. In someembodiments, one, two, three, four, or all five of the above may beintegrated in a single detection station. The single detections stationmay be configured to simultaneously measure multiple different assays atthe same time.

The detection unit may be above the component from which the signal isdetected, beneath the component from which the signal is detected, tothe side of the component from which the signal is detected, orintegrated into the component from which the signal is detected, or mayhave different orientation in relation to the component from which thesignal is detected. For example, the detection unit may be incommunication with an assay unit. The detection unit may be proximate tothe component from which the signal is detected, or may be remote to thecomponent from which the signal is detected. The detection unit may bewithin one or more mm, one or more cm, one or more 10s of cm from whichthe component from which the signal is detected.

The detection unit may have a fixed position, or may be movable. Thedetection unit may be movable relative to a component from which asignal is to be detected. For example, the detection unit can be movedinto communication with an assay unit or the assay unit can be movedinto communication with the detection unit. In one example, a sensor isprovided to locate an assay unit relative to a detector when an assay isdetected.

A detection unit may include one or more optical or visual sensor orsonic or magnetic or radioactivity sensor or some combination of these.For example, a detection unit may include microscopy, visual inspection,via photographic film, or may include the use of electronic detectorssuch as digital cameras, charge coupled devices (CCDs), super-cooled CCDarrays, photodetector or other detection device. An optical detector mayfurther include non-limiting examples include a photodiode,photomultiplier tube (PMT), photon counting detector, or avalanche photodiode, avalanche photodiode arrays. In some embodiments a pin diode maybe used. In some embodiments a pin diode can be coupled to an amplifierto create a detection device with a sensitivity comparable to a PMT.Some assays may generate luminescence as described herein. In someembodiments fluorescence or chemiluminescence is detected. In someembodiments a detection assembly could include a plurality of fiberoptic cables connected as a bundle to a CCD detector or to a PMT array.The fiber optic bundle could be constructed of discrete fibers or ofmany small fibers fused together to form a solid bundle. Such solidbundles are commercially available and easily interfaced to CCDdetectors. In some embodiments, fiber optic cables may be directlyincorporated into assay or reagent units. For example, samples or tipsas described elsewhere herein may incorporate fiber optic cables. Insome embodiments, electronic sensors for detection or analysis (such asimage processing) may be built into the pipette or other component ofthe fluid handling system. In some embodiments, a detection unit may bea PMT. In some embodiments, a detection unit may be a photodiode. Insome embodiments, a detection unit may be a spectrophotometer. In someembodiments, a detection unit may be a nucleic acid assay stationcontaining or operatively coupled to a light source and an opticalsensor. In some embodiments, a detection unit may be a camera. In someembodiments, a detection unit may be an imaging device. In someembodiments, a detection unit may be a cytometry station containing amicroscopy stage and an imaging device. In some embodiments, a detectionunit containing a CCD or CMOS sensor may be configured to obtain adigital image, such as of a sample, assay unit, cuvette, assay, thedevice, or the device surroundings. The digital image may betwo-dimensional or three-dimensional. The digital image may be a singleimage or a collection of images, including video. In some instances,digital imaging may be used by the device or system for control ormonitoring of the device, it surroundings, or processes within thedevice.

One or more detection units may be configured to detect a detectablesignal, which can be a light signal, including but not limited tophotoluminescence, electroluminescence, sonoluminescence,chemiluminescence, fluorescence, phosphorescence, polarization,absorbance, turbidity or scattering. In some embodiments, one or morelabel may be employed during a chemical reaction. The label may permitthe generation of a detectable signal. Methods of detecting labels arewell known to those of skill in the art. Thus, for example, where thelabel is a radioactive label, means for detection may include ascintillation counter or photographic film as in autoradiography. Wherethe label is a fluorescent label, it may be detected by exciting thefluorochrome with the appropriate wavelength of light and detecting theresulting fluorescence by, for example, microscopy, visual inspection,via photographic film, by the use of electronic detectors such asdigital cameras, charge coupled devices (CCDs) or photomultipliers andphototubes, or other detection device. In some embodiments, imagingdevices may be used, such as cameras. In some instances, cameras may useCCDs, CMOS, may be lensless cameras (e.g., Frankencamera),microlens-array cameras, open-source cameras, or may use or any othervisual detection technology known or later developed in the art. Camerasmay acquire non-conventional images, e.g. holographic images,tomographic or interferometric, Fourier-transformed spectra, which maythen be interpreted with or without the aid of computational methods.Cameras may include one or more feature that may focus the camera duringuse, or may capture images that can be later focused. In someembodiments, imaging devices may employ 2-d imaging, 3-d imaging, and/or4-d imaging (incorporating changes over time). Imaging devices maycapture static images. The optical schemes used to achieve 3-D and 4-Dimaging may be one or more of the several known to those skilled in theart, e.g. structured illumination microscopy (SLM), digital holographicmicroscopy (DHM), confocal microscopy, light field microscopy etc. Thestatic images may be captured at one or more point in time. The imagingdevices may also capture video and/or dynamic images. The video imagesmay be captured continuously over one or more periods of time. Animaging device may collect signal from an optical system which scans thesample in arbitrary scan patterns (e.g. raster scan). In someembodiments, the imaging device may use one or more component of thedevice in capturing the image. For example, the imaging device may use atip and/or vessel to assist with capturing the image. The tip and/orvessel may function as an optic to assist in capturing an image.

Detection units may also be capable of capturing audio signals. Theaudio signals may be captured in conjunction with one or more image.Audio signals may be captured and/or associated with one or more staticimage or video images. Alternatively, the audio signals may be capturedseparate from the image.

In one example, a PMT may be used as a detector. In some instances,count rates as low as 100 per second and count rates as high as10,000,000 may be measurable. The linear response range of PMTs (forexample, the range where count rate is directly proportional to numberof photons per unit time) can be about 1000-3,000,000 counts per second.In an example, an assay has a detectable signal on the low end of about200-1000 counts per second and on the high end of about 10,000-2,000,000counts per second. In some instances for protein biomarkers, the countrate is directly proportional to alkaline phosphatase bound to thecapture surface and also directly proportional to the analyteconcentration.

In another example, a detector may include a camera that may be imagingin real-time. Alternatively, the camera may take snapshots at selectedtime intervals or when triggered by an event. Similarly, the camera maytake video at selected time intervals or when triggered by an event. Insome embodiments, the camera may image a plurality of samplessimultaneously. Alternatively, the camera may image a selected view, andthen move on to a next location for a different selected view.

A detection unit may have an output that is digital and generally aone-to-one or one-to-many transformation of the detected signal, e.g.,the image intensity value is an integer proportional to a positive powerof the number of photons reaching the camera sensor over the time ofexposure. Alternatively, the detection unit may output an analog signal.The detectable range for exemplary detectors can be suitable to thedetector being used.

The detection unit may be capable of capturing and/or imaging a signalfrom anywhere along the electromagnetic spectrum. For example, adetection unit may be capable of capturing and/or imaging visiblesignals, infra-red signals, near infra-red signals, far infra-redsignals, ultraviolet signals, gamma rays, microwaves, and/or othersignals. The detection unit may be capable of capturing acoustic wavesover a large range of frequencies, e.g. audio, ultrasound. The detectionunit may be capable of measuring magnetic fields with a wide range ofmagnitude.

An optical detector can also comprise a light source, such as anelectric bulb, incandescent bulb, electroluminescent lamp, laser, laserdiode, light emitting diode (LED), gas discharge lamp, high-intensitydischarge lamp, natural sunlight, chemiluminescent light sources. Otherexamples of light sources as provided elsewhere herein. The light sourcecan illuminate a component in order to assist with detecting theresults. For example, the light source can illuminate an assay in orderto detect the results. For example, the assay can be a fluorescenceassay or an absorbance assay, as are commonly used with nucleic acidassays. The detector can also comprise optics to deliver the lightsource to the assay, such as a lens, mirror, scanning or galvano-mirror,prisms, fiber optics, or liquid light guides. The detector can alsocomprise optics to deliver light from an assay to a detection unit. Insome embodiments, a light source can be coupled to an opticaldetector/sensor which is configured primarily for the detection ofluminescent assays, in order to expand the range of types of assays thatmay be detected by the optical sensor (e.g. to include absorbance,fluorescence, turbidity, and colorimetry assays, etc.).

An optical detection unit may be used to detect one or more opticalsignal. For example, the detection unit may be used to detect a reactionproviding luminescence. The detection unit may be used to detect areaction providing fluorescence, chemiluminscence, photoluminescence,electroluminescence, color change, sonoluminescence, absorbance,turbidity, or polarization. The detection unit may be able to detectoptical signals relating to color intensity and phase or spatial ortemporal gradients thereof. For example, the detection unit may beconfigured to detect selected wavelengths or ranges of wavelengths. Theoptical detection unit may be configured to move over the sample and amirror could be used to scan the sample simultaneously.

In some embodiments, an assay provided herein generating a particulartype of result (e.g. luminescence, turbidity, color change/colorimetry,etc.) may be monitored by different types or configurations of detectionunits provided herein. For example, in some situations, an assayresulting in a turbid reaction product may be monitored in: i) adedicated spectrophotometer, ii) a nucleic acid assay station containingor operatively coupled to a light source and a optical sensor, or iii) adetection unit containing a CCD sensor (e.g. a stand-alone imagingdevice containing a CCD sensor, or a cytometry station containing animaging device containing a CCD sensor). In both detection unitconfigurations i) and ii), the sample may be positioned in the detectionunit between the respective light source and the respective opticalsensor, such that I₀ (incident radiation) and I₁ (transmitted radiation)values may be measured at one or more selected wavelengths, andabsorbance calculated. In detection unit configuration iii), an image ofthe sample may be obtained by the CCD sensor, and further processed byimage analysis. In some embodiments, a sample may be monitored in morethan one of the above detection units. In another example, in somesituations, an assay resulting in a chemiluminescent signal may bemonitored by i) a photodiode or other luminescence sensor, ii) a nucleicacid assay station containing or operatively coupled to a light sourceand an optical sensor, or iii) a detection unit containing a CCD sensor.In configuration i) the photodiode detects light from thechemiluminescent reaction. In some situations, the photodiode may beconfigured to sense very low levels of light, and thus may be used withassays which result in only a low level of chemiluminescence. Inconfiguration ii) the assay (including non-nucleic acid amplificationassays) may be placed in the nucleic acid amplification module, and theoptical sensor within the station may be used to detect light from thechemiluminescent assay (without using the light source in the station).In some situations, the optical sensor in this configuration may not beas sensitive to light as a stand-alone photodiode or PMT, and therefore,use of the nucleic acid assay station as detector for chemiluminscenceassays may be with assays which produce relatively moderate to highlevels of chemiluminescent light. In configuration iii), an image of thechemiluminescent sample may be obtained by the CCD sensor, and furtherprocessed by image analysis (including light counts) to determine thelevel of chemiluminescence in the sample.

In some embodiments, the controller of a system, device, or moduleprovided herein may be configured to select a particular detection unitfrom two or more detection units within a device or module for thedetection of a signal or data from a selected assay unit within the samedevice or module. For example, a module of a device provided herein maycontain three detection units: i) a photodiode, ii) a nucleic acid assaystation containing or operatively coupled to a light source and anoptical sensor, and iii) a detection unit containing a CCD sensor. Themodule may also contain multiple assay units and may simultaneouslyperform multiple assays. Before, during, or after the performance of,for example, a chemiluminescent assay in a particular movable assay unitin an assay station in the module, the controller may determine which ofthe three detection units in the module to use for receiving theselected assay unit and detecting a signal or data from the assay unit.In making the determination, the controller may take into account one ormore factors, such as: i) detection unit availability—one or more of thedetection units may be occupied with other assay units at the time ofthe completion of the assay in the selected assay unit; ii) detectionunit suitability for receiving a particular assay unitconfiguration—different detection units may be optimized for receivingassay units of particular shapes or sizes; iii) detection unitsuitability for detecting the signal or data from the particular assaybeing performed within the selected assay unit—different detection unitsmay be optimized to measure a particular property of a sample (e.g.absorbance vs. fluorescence vs. color, etc.), or different detectionunits may be optimized to measure certain features/versions of aparticular property of a sample (e.g. a detection unit containing anoptical sensor may be optimized to measure high levels of light or lowlevels of light, or a detection unit configured for measuringfluorescence may be configured to measure the fluorescence of compoundshaving a certain range of excitation wavelengths and a certain range ofemission wavelengths); and iv) total time for multiplexing of assays—inorder to reduce the total time necessary to perform or obtain data frommultiple assays within the device or module, the controller may takeinto account other assays simultaneously being performed in the deviceor module, such that the use of each detection unit is optimized for thecombination of all assays being simultaneously performed in the moduleor device. Based on the various determinations by the controller, thecontroller may direct a sample handling apparatus (for example, apipette) within the module to transport the assay unit containing thechemiluminescent assay to a particular detection unit within the module,for measurement of the chemiluminescent signal. In this example, if thechemiluminescent assay in the selected assay unit is expected togenerate a low level of light and the photodiode is available at thetime of the completion of the assay in the selected assay unit, thecontroller may direct the sample handling apparatus to transport theselected assay unit to the photodiode for measurement. In someembodiments, the controller may contain a protocol for the detection ofan assay in a selected assay unit with a detection unit selected fromtwo or more detection units in the module or device, where the protocoltakes into account one or more of the factors indicated above relevantto the selection of a detection unit from two or more detection units.The protocol may be stored in the module or the device, stored in anexternal device or cloud, or generated on demand. Protocols that aregenerated on demand may be generated on the device or on an externaldevice or cloud, and downloaded to the sample processing device.

In some embodiments, the device or controller may receive or store aprotocol which contains instructions for directing a sample handlingapparatus within a device or module to move assay units to differentdetection units (or vice versa) in the device or module, and which takesinto account multiple assays being simultaneously performed in the samemodule or device. Optionally, with such protocols, different assayshaving the same reaction outcome may be measured in different detectionunits provided herein (e.g. a chemiluminescent reaction may be measuredin, for example, a PMT or a camera containing a CCD sensor), dependingon the other assays being performed simultaneously in the same module ordevice. These features of the controller, protocols, and detection unitsprovide multiple benefits, including, for example, the ability toefficiently multiplex discrete assays within a device or module, and theability to efficiently obtain data from assays using different detectionunits.

In some embodiments, the detection system may comprise optical ornon-optical detectors or sensors for detecting a particular parameter ofa subject. Such sensors may include sensors for temperature, electricalsignals, for compounds that are oxidized or reduced, for example, O₂,H₂O₂, and I₂, or oxidizable/reducible organic compounds. Detectionsystem may include sensors which measure acoustic waves, changes inacoustic pressure and acoustic velocity. In some embodiments, systemsand devices provided herein may contain a barometer or other device forsensing atmospheric pressure. Atmospheric pressure measurements may beuseful, for example, for adjusting protocols to high or low-pressuresituations. For example, atmospheric pressure may be relevant to assaysthat measure one or more dissolved gases in a sample. In addition,atmospheric pressure measurements may be useful, for example, when usinga device provided herein in high or low pressure environment (e.g. athigh altitudes, on an airplane, or in space).

Examples of temperature sensors may include thermometers, thermocouples,or IR sensors. The temperature sensors may or may not use thermalimaging. The temperature sensor may or may not contact the item whosetemperature is to be sensed.

Examples of sensors for electrical properties may include sensors thatcan detect or measure voltage level, current level, conductivity,impedance, or resistance. Electrical property sensors may also includepotentiometers or amperometric sensors.

In some embodiments, labels may be selected to be detectable by adetection unit. The labels may be selected to be selectively detected bya detection unit. Examples of labels are discussed in greater detailelsewhere herein.

Any of the sensors may be triggered according to one or more schedule,or a detected event. In some embodiments, a sensor may be triggered whenit receives instructions from one or more controller. A sensor may becontinuously sensing and may indicate when a condition is sensed.

The one or more sensors may provide signals indicative of measuredproperties to a controller. The one or more sensors may provide signalsto the same controller or to different controllers. In some embodiments,the controller may have a hardware and/or software module which mayprocess the sensor signal to interpret it for the controller. In someembodiments, the signals may be provided to the controller via a wiredconnection, or may be provided wirelessly. The controller may beprovided on a system-wide level, group of device level, device level,module level, or component of module level, or any other level asdescribed elsewhere herein.

The controller may, based on the signals from the sensors, effect achange in a component or maintain the state of a unit. For example, thecontroller may change the temperature of a thermal control unit, modifythe rotation speed of a centrifuge, determine a protocol to run on aparticular assay sample, move a vessel and/or tip, or dispense and/oraspirate a sample. In some embodiments, based on the signals from thesensors, the controller may maintain one or more condition of thedevice. One or more signal from the sensors may also permit thecontroller to determine the current state of the device and track whatactions have occurred, or are in progress. This may or may not affectthe future actions to be performed by the device. In some instances, thesensors (e.g., cameras) may be useful for detecting conditions that mayinclude errors or malfunctions of the device. The sensors may detectconditions that may lead to an error or malfunction in data collection.Sensors may be useful in providing feedback in trying to correct adetected error or malfunction.

In some embodiments, one or more signal from a single sensor may beconsidered for particular actions or conditions of the device.Alternatively, one or more signals from a plurality of sensors may beconsidered for particular actions or conditions of the device. The oneor more signals may be assessed based on the moment they are provided.Alternatively, the one or more signals may be assessed based oninformation collected over time. In some embodiments, the controller mayhave a hardware and/or software module which may process one more sensorsignals in a mutually-dependent or independent manner to interpret thesignals for the controller.

In some embodiments, multiple types of sensors or detection units may beuseful for measuring the same property. In some instances, multipletypes of sensors or detection units may be used for measuring the sameproperty and may provide a way of verifying a measured property or as acoarse first measurement which can then be used to refine the secondmeasurement. For example, both a camera and a spectroscope or other typeof sensor may be used to provide a colorimetric readout. Nucleic acidassay may be viewed via fluorescence and another type of sensor. Cellconcentration can be measured with low sensitivity using absorbance orfluorescence with the aim of configuring the same or another detectorprior to performing high sensitivity cytometry. With systems, devices,methods, and assays provided herein, turbidity of a sample may beassayed, for example, by measuring i) the light transmitted through thesample (similar to an absorbance measurement and may includecolorimetry; the light path may travel through the sample horizontallyor vertically); or ii) the light scattered by the sample (sometimesknown as a nephelometric measurement). Typically, for option i), thelight sensor is located in-line with the light source, and the sample tobe measured is located between the light source and the optical sensor.Typically, for option ii), the optical sensor is off-set from path ofthe light from the light source (e.g. at a 90 degree angle), and thesample to be measured is located in the path of the light source. Inanother example, agglutination of a sample may be assayed, for example,by: i) measuring the light transmitted through the sample (similar to anabsorbance measurement and may include colorimetry); ii) measuring lightscattered by the sample (sometimes known as a nephelometricmeasurement); iii) obtaining an electronic image of the sample (e.g.with a CCD or CMOS optical sensor), followed by manual or automatedimage analysis; or iv) visual inspection of the sample.

The controller may also provide information to an external device. Forexample, the controller may provide an assay reading to an externaldevice which may further analyze the results. The controller may providethe signals provided by the sensors to the external device. Thecontroller may pass on such data as raw data as collected from thesensors. Alternatively, the controller may process and/or pre-processthe signals from the sensors before providing them to the externaldevice. The controller may or may not perform any analysis on thesignals received from the sensors. In one example the controller may putthe signals into a desired format without performing any analysis.

In some embodiments, detection units may be provided inside a housing ofthe device. In some instances, one or more detection units, such assensors may be provided external to the housing of the device. In someembodiments, a device may be capable of imaging externally. For example,the device may be capable of performing MRI, ultrasound, or other scans.This may or may not utilize sensors external to the device. In someinstances, it may utilize peripherals, which may communicate with thedevice. In one example a peripheral may be an ultrasonic scanner. Theperipherals may communicate with the device through a wireless and/orwired connection. The device and/or peripherals may be brought intoclose proximity (e.g., within 1 m, 0.5 m, 0.3 m, 0.2 m, 0.1 cm, 8 cm, 6cm, 5 cm, 4 cm, 3 cm, 2 cm, 1 cm, 0.5 cm) or contact the area to bescanned. In some embodiments, a device may contain or communicate with aperipheral device for performing x-rays (e.g. x-ray generator anddetector), sonography, ultrasound, or echocardiograms (e.g. sonographicscanners), a cooximeter, or eye scans (e.g. optical sensor). In someembodiments, a device may contain or communicate with an independentlymovable peripheral that can, with aid of an imaging device, physicallyfollow a subject (e.g. throughout a room or a house), and monitor thesubject. The independently movable peripheral may, for example, monitorsubjects that require a high level of care or monitoring.

In some embodiments, a sensor may be integrated into a pill or patch. Insome embodiments, a sensor may be implantable or injectable. Optionally,such a sensor may be a multi-analyte sensor that is implanted/injected.All such sensors (pill, patch, implanted/injected) could measure themultiple assay methodologies simultaneously, sequentially, or singly andmay communicate with a cell phone or external device by way of wired,wireless, or other communication technique. Any of these sensors may beconfigured to performed one or more types of assays or obtain one ormore types of data from a subject (e.g. temperature, electrochemical,etc.). Data from the sensors may be, for example, communicated to anexternal device or a sample processing device of a system providedherein. In some embodiments, the sensors may receive instructions froman external device or a sample processing device regarding, for example,when to perform a measurement or what assay to perform.

Cameras

Cameras described herein may be charge coupled device (CCDs) cameras,super-cooled CCD cameras, or other optical cameras. Such cameras may beformed on chips having one or more cameras, such as part of an array ofcameras. Such cameras may include one or more optical components, forexample, for capturing light, focusing light, polarizing light,rejecting unwanted light, minimizing scattering, improving imagequality, improving signal-to-noise. In an example, cameras may includeone or more of lenses and mirrors. Such cameras may have color ormonochromatic sensors. Such cameras may also include electroniccomponents such as microprocessors and digital signal processors for oneor more of the following tasks: image compression, improvement ofdynamic range using computational methods, auto-exposure, automaticdetermination of optimal camera parameters, image processing, triggeringstrobe light to be in sync with the camera, in-line controller tocompensate for effect of temperature changes on camera sensorperformance. Such cameras may also include on-board memory to bufferimages acquired at high frame rates. Such cameras may include mechanicalfeatures for image quality improvement such as a cooling system oranti-vibration system.

Cameras may be provided at various locations of point of servicesystems, devices and modules described herein. In an embodiment, camerasare provided in modules for imaging various processing routines,including sample preparation and assaying. This may enable the system todetect a fault, perform quality control assessments, performlongitudinal analysis, perform process optimization and synchronizeoperation with other modules and/or systems.

In some cases, a camera includes one or more optical elements selectedfrom the group consisting of a lens, a mirror, a diffraction grating, aprism, and other components for directing and/or manipulating light. Inother cases, a camera is a lens-less (or lensless) camera configured tooperate without one or more lenses. An example of a lens-less camera isthe Frankencamera. In an embodiment, a lens-less camera uses (orcollects) reflected or scattered light and computer processing to deducethe structure of an object.

In an embodiment, a lens-less camera has a diameter of at most about 10nanometers (“nm”), at most about 100 nm, at most about 1 μm, at mostabout 10 μm, at most about 100 μm, at most about 1 mm, at most about 10mm, at most about 100 mm, or at most about 500 mm. In anotherembodiment, a lens-less camera has a diameter between about 10 nm and 1mm, or between about 50 nm and 500 μm.

Cameras provided herein are configured for rapid image capture. Systememploying such cameras may provide images in a delayed fashion, in whichthere is a delay from the point in which an image is captured to thepoint it is displayed to a user, or in real-time, in which there is lowor no delay from the point in which an image is captured to the point itis displayed to the user. In some situations, cameras provided hereinare configured to operate under low or substantially low lightingconditions.

In some situations, cameras provided herein are formed of opticalwaveguides configured to guide electromagnetic waves in the opticalspectrum. Such optical waveguides may be formed in an array of opticalwaveguides. An optical waveguide may be a planar waveguide, which mayinclude one or more gratings for directing light. In some cases, thecamera may have fiber optic image bundles, image conduits or faceplatescarrying light to the camera sensor.

Cameras may be useful as detection units. Cameras may also be useful forimaging one or more sample or portion of a sample. Cameras may be usefulfor pathology. Cameras may also be useful for detecting theconcentration of one or more analyte in a sample. Cameras may be usefulfor imaging movement or change of a sample and/or analytes in a sampleover time. Cameras may include video cameras that may capture imagescontinuously. Cameras may also optionally capture images at one or moretimes (e.g., periodically, at predetermined intervals (regular orirregular intervals), in response to one or more detected event). Forexample, cameras may be useful for capturing changes of cell morphology,concentration and spatial distribution of entities in cells that arelabeled with contrast agents (e.g. fluorescent dyes, gold nanoparticles)and/or movement. Cell imaging may include images captured over time,which may be useful for analyzing cell movement and morphology changes,and associated disease states or other conditions. Cameras may be usefulfor capturing sample kinematics, dynamics, morphology, or histology.Such images may be useful for diagnosis, prognosis, and/or treatment ofa subject. An imaging device may be a camera or a sensor that detectsand/or records electromagnetic radiation and associated spatial and/ortemporal dimensions.

Cameras may be useful for interaction of an operator of a device withthe device. The cameras may be used for communications between a deviceoperator and another individual. The cameras may permit teleconferencingand/or video conferencing. The cameras may permit a semblance offace-to-face interactions between individuals who may be at differentlocations. Images of a sample or component thereof, or an assay orreaction involving same, may be stored, enabling subsequent reflextesting, analysis and/or review. Image processing algorithms may be usedto analyze collected images within the device or remotely.

Cameras may also be useful for biometric measurements (e.g., waistcircumference, neck circumference, arm circumference, leg circumference,height, weight, body fat, BMI) of a subject and/or identifying a subjector operator of a device (e.g., facial recognition, retinal scan,fingerprint, handprint, gait, movement) which may optionally becharacterized through imaging. Embedded imaging systems may also captureultrasound or MRI (magnetic resonance imaging) of a subject through thesystem. Cameras may also be useful for security applications, asdescribed elsewhere herein. Cameras may also be useful for imaging oneor more portion of the device and for detecting error within the device.Cameras may image and/or detect a malfunction and/or proper function ofmechanics of one or more component of the device. Cameras may be used tocapture problems, correct a problem, or learn from detected conditions.For example, a camera may detect whether there is an air bubble in thetip, which may end up skewing readouts or may result in error. A cameramay also be used to detect if a tip is not properly bound to a pipette.Cameras may capture images of components and determine whether thecomponents are positioned properly, or where components are positioned.Cameras may be used as part of a feedback loop with a controller todetermine the location of components with sub-micrometer resolution andadjust system configuration to account for the exact location.

Dynamic-Resource Sharing

One or more resource of a device may be shared. Resource-sharing mayoccur at any level of the device. For example, one or more resource of amodule may be shared within the module. In another example, one or moreresource of a device may be shared between modules. One or more resourceof a rack may be shared within a rack. One or more resource of a devicemay be shared between racks.

A resource may include any component of a device, reagent providedwithin a device, sample within the device, or any other fluid within thedevice. Examples of components may include but are not limited to fluidhandling mechanism, tip, vessel, assay unit, reagent unit, dilutionunit, wash unit, contamination reduction mechanism, filter, centrifuge,magnetic separator, incubator, heater, thermal block, cytometer, lightsource, detector, housing, controller, display, power source,communication unit, identifier, or any other component known in the artor described elsewhere herein. Other examples of components may includereagents, wash, diluents, sample, labels, or any fluid or substance thatmay be useful for effecting a chemical reaction. A module may include,one, two, three, four, five, or more of the resources listed herein. Adevice may include one, two, three, four, five, or more of the resourceslisted herein. The modules may include different resources, or mayinclude the same resources. A device may include one or more modules notprovided within a module.

It may be desirable to use a resource that may not be readily available.A resource may be not readily available when the resource is being used,is scheduled to be used, does not exist, or is inoperable. For example,within a module it may be desirable to centrifuge a sample, while themodule may not have a centrifuge, the centrifuge may be in use, and/orthe centrifuge may be undergoing an error. The device may determinewhether an additional centrifuge is available within the module. If anadditional centrifuge is available within the module, then the devicemay use the available centrifuge. This may apply to any resource withinthe module. In some embodiments, a resource within the module may beable to compensate for a deficiency in another. For example, if twocentrifuges are needed, but one is out of commission, the othercentrifuge may be used to accommodate both centrifugationssimultaneously, or in sequence.

In some instances, the desired resource may not be available within theselected module, but may be available in another module. The resource inthe other module may be used. For example, if a centrifuge in a firstmodule breaks, is in use, or does not exist, a centrifuge in a secondmodule may be used. In some embodiments, a sample and/or other fluid maybe transferred from the first module to the second module to use theresource. For example, a sample may be transferred from the first moduleto the second module to use the centrifuge. Once the resource has beenused, the sample and/or other fluid may be transferred back to the firstmodule, may remain at the second module, or may be transferred to athird module. For example, the sample may be transferred back to thefirst module for further processing, using resources available in thefirst module. In another example, the same may remain in the secondmodule for further processing, if needed resources are available in thesecond module. In another example, if the resources needed are notavailable in the first and second module, or the scheduling is somehowimproved by using a resource at a third module, the sample and/or otherfluid may be transferred to the third module.

The sample and/or other fluids may be transferred between modules. Insome embodiments, a robotic arm may shuttle a sample, reagent, and/orother fluids between modules, as described in greater detail elsewhereherein. The sample and/or other fluids may be transferred using a fluidhandling system. The sample and/or other fluids may be transferredbetween modules within tips, vessels, units, compartments, chambers,tubes, conduits, or any other fluid containing and/or transferringmechanisms. In some embodiments, fluid may be contained withinfluidically isolated or hydraulically independent containers while beingtransferred between modules. Alternatively, they may flow through aconduit between modules. The conduits may provide fluid communicationbetween modules. Each module may have a fluid handling system ormechanism that may be able to control the movement of the sample and/orfluid within the module. A first fluid handling mechanism in the firstmodule may provide the fluid to an inter-module fluid transport system.A second fluid handling mechanism at a second module may pick up thefluid from the inter-module fluid transport system and may transfer thefluid in order to enable the use of a resource in the second module.

In alternate embodiments, one or more resource may be transferredbetween modules. For example, a robotic arm may shuttle a resourcebetween modules. Other mechanisms may be used to transfer a resourcefrom a first module to a second module. In one example, a first modulemay contain a reagent within a reagent unit. The reagent and reagentunit may be transferred to the second module which may use the reagentand reagent unit.

A resource may be provided within a device that may be external to allmodules. A sample and/or other fluid may be transferred to thisresource, and the resource may be used. The sample and/or fluid may betransferred to the resource external to the modules using a robotic armor any other transferring mechanism described elsewhere herein.Alternatively, the external resource may be transferred to one or moremodule. In one example, a cytometer may be provided within a device, butexternal to all modules. In order to access the cytometer, samples maybe shuttled to and from modules to the cytometer.

Such allocations of resources within modules, between modules, or withinthe device external to modules may occur dynamically. The device may becapable of tracking which resources are available. Based on one or moreprotocol, the device may be able to determine on the fly whether aresource is available or unavailable. The device may also be able todetermine whether another of the resource is available within the samemodule, different module, or elsewhere within the device. The device maydetermine whether to wait to use a currently unavailable resource, or touse another available resource depending on one or more set ofprotocols. The device may be able to track whether a resource willbecome unavailable in the future. For example, a centrifuge may bescheduled to be used after a sample has been incubated a predeterminedlength of time. The centrifuge may be unavailable starting from the timeof intended use to the anticipated end of use. The future unavailable ofa resource may be accounted for by a protocol.

In some embodiments, signals from one or more sensors may assist withthe on-the-fly determination on the status of a resource and/or theavailability of the resource. One or more sensors and/or the detectormay be able to provide real-time feedback or updates on the status of aresource and/or process. The system may determine whether adjustmentsneed to be made to a schedule and/or whether the use another resource.

A protocol may include one or more set of instructions that maydetermine which resources to use at which times. The protocol mayinclude instructions to use resources within the same module, withindifferent modules, or external to the module. In some embodiments, theprotocol may include one or more set of priorities or criteria. Forexample, if a resource within the same module is available, this may beused rather than a module that is provided within another module. Aresource that is in closer proximity to the sample using the resourcemay have a higher priority. For example, if one or more step is beingperformed on a sample within a first module, and the resource isavailable within the first module, then the resource may be used. Ifmultiple copies of the resource are available within the first module,the copy of the resource closest to the sample may be used. If theresource is unavailable within the first module, the resource availablein the closest module to the first module may be used. In anotherexample, current and future availability may also be taken into accountfor determining the use of a module. This information may come from theCloud, the controller, the device or from the module itself. In someembodiments, speed of completion may be prioritized higher thanproximity (e.g., trying to keep samples within the same module).Alternatively, proximity may be prioritized higher than speed. Othercriteria may include but are not limited to, proximity, speed, time ofcompletion, fewer steps, or less amount of energy consumed. The criteriamay have any ranking in order of preference, or any other set ofinstructions or protocols may determine the use of resources and/orscheduling.

Housing

In accordance with some embodiments of the invention, a system mayinclude one or more devices. A device may have a housing and/or supportstructure.

In some embodiments, a device housing may entirely enclose the device.In other embodiments, the device housing may partially enclose thedevice. The device housing may include one, two, three, four, five, sixor more walls that may at least partially enclose the device. The devicehousing may include a bottom and/or top. The device housing may containone or more modules of the device within the housing. The device housingmay contain electronic and/or mechanical components within the housing.The device housing may contain a fluid handling system within thehousing. The device housing may contain one or more communication unitwithin the housing. The device housing may contain one or morecontroller unit. A device user interface and/or display may be containedwithin the housing or may be disposed on a surface of the housing. Adevice may or may not contain a power source, or an interface with apower source. The power source may be provided or interfaced within thehousing, external to the housing, or incorporated within the housing.

A device may or may not be air tight or fluid tight. A device may or maynot prevent light or other electromagnetic waves from entering thehousing from outside the device, or escaping the housing from within thedevice. In some instances, individual modules may or may not be airtight or fluid tight and/or may or may not prevent light or otherelectromagnetic waves from entering the module.

In some embodiments, the device may be supported by a support structure.In some embodiments, the support structure may be a device housing. Inother embodiments, a support structure may support a device from beneaththe device. Alternatively, the support structure may support a devicefrom one or more side, or from the top. The support structure may beintegrated within the device or between portions of the device. Thesupport structure may connect portions of the device. Any description ofthe device housing herein may also apply to any other support structureor vice versa.

The device housing may fully or partially enclose the entire device. Thedevice housing may enclose a total volume of less than or equal to about4 m³, 3 m³, 2.5 m³, 2 m³, 1.5 m³, 1 m³, 0.75 m³, 0.5 m³, 0.3 m³, 0.2 m³,0.1 m³, 0.08 m³, 0.05 m³, 0.03 m³, 0.01 m³, 0.005 m³, 0.001 m³, 500 cm³,100 cm³, 50 cm³, 10 cm³, 5 cm³, 1 cm³, 0.5 cm³, 0.1 cm³, 0.05 cm³, or0.01 cm³. The device may have any of the volumes described elsewhereherein.

The device and/or device housing may have a footprint covering a lateralarea of the device. In some embodiments, the device footprint may beless than or equal to about 4 m², 3 m², 2.5 m², 2 m², 1.5 m², 1 m², 0.75m², 0.5 m², 0.3 m², 0.2 m², 0.1 m², 0.08 m², 0.05 m², 0.03 m², 100 cm²,80 cm², 70 cm², 60 cm², 50 cm, 40 cm², 30 cm², 20 cm², 15 cm², 10 cm², 7cm², 5 cm², 1 cm², 0.5 cm², 0.1 cm², 0.05 cm, or 0.01 cm².

The device and/or device housing may have a lateral dimension (e.g.,width, length, or diameter) or a height less than or equal to about 4 m,3 m, 2.5 m, 2 m, 1.5 m, 1.2 m, 1 m, 80 cm, 70 cm, 60 cm, 50 cm, 40 cm,30 cm, 25 cm, 20 cm, 15 cm, 12 cm, 10 cm, 8 cm, 5 cm, 3 cm, 2 cm, 1 cm,0.5 cm, 0.1 cm, 0.05 cm, or 0.01 cm. The lateral dimensions and/orheight may vary from one another. Alternatively, they may be the same.In some instances, the device may be a tall and thin device, or may be ashort and squat device. The height to lateral dimension ratio may begreater than or equal to 100:1, 50:1, 30:1, 20:1, 10:1, 9:1, 8:1, 7:1,6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9,1:10, 1:20, 1:30, 1:50, or 1:100.

The device and/or device housing may have any shape. In someembodiments, the device may have a lateral cross-sectional shape of arectangle or square. In other embodiments, the device may have a lateralcross-sectional shape of a circle, ellipse, triangle, trapezoid,parallelogram, pentagon, hexagon, octagon, or any other shape. Thedevice may have a vertical cross-sectional shape of a circle, ellipse,triangle, rectangle, square, trapezoid, parallelogram, pentagon,hexagon, octagon, or any other shape. The device may or may not have abox-like shape. The device may or may not have a flattened planar shapeand/or a rounded shape.

A device housing and/or support structure may be formed of a rigid,semi-rigid or flexible material. A device housing may be formed of oneor more materials. In some embodiments, the device housing may includepolystyrene, moldable or machinable plastic. The device housing mayinclude polymeric materials. Non-limiting examples of polymericmaterials include polystyrene, polycarbonate, polypropylene,polydimethysiloxanes (PDMS), polyurethane, polyvinylchloride (PVC),polysulfone, polymethylmethacrylate (PMMA),acrylonitrile-butadiene-styrene (ABS), and glass. The device housing maybe an opaque material, a translucent material, a transparent material,or may include portions that are any combination thereof.

The device housing may be formed of a single integral piece or multiplepieces. The device housing may comprise multiple pieces that may bepermanently affixed to one another or removably attached to one another.In some instances, one or more connecting features of the housing may becontained within the housing only. Alternatively one or more connectingfeatures of the device housing may be external to the device housing.The device housing may be opaque. The device housing may preventuncontrolled light from entering the device. The device housing mayinclude one or more transparent portions. The device housing may permitcontrolled light to enter selected regions of the device.

The device housing may contain one or more movable portion that may beused to accept a sample into the device. Alternatively, the devicehousing may be static as a sample is provided to the device. For examplethe device housing may include an opening. The device opening may remainopen or may be closable. A device opening may directly or indirectlylead to a sample collection unit, such that a subject may provide asample to the device through the device housing. In such circumstances,the sample may be provided, for example, to a cartridge in the device.The device may include one or more movable tray that may accept one ormore sample or other component of the device. The tray may betranslatable in a horizontal and/or vertical direction. The opening maybe in fluid communication with one or more portion of the fluid handlingsystem therein. The opening may be selectively opened and/or closed. Oneor more portions of the device housing may be selectively opened and/orclosed.

In some embodiments, the device housing may be configured to accept acartridge, or sample collection unit. In some embodiments, the devicehousing may be configured to accept or collect a sample. The devicehousing may be configured to collect a sample directly from a subject oran environment. The sample receiving location may be configured to havean opened and a closed position, such that when closed, the devicehousing may be sealed. The device housing may be in contact with thesubject or environment. Additional details relating to sample collectionmay be described elsewhere herein.

In some embodiments, the housing may surround one or more of the racks,modules, and/or components described elsewhere herein. Alternatively,the housing may be integrally forming one or more of the racks, modules,and/or components described elsewhere herein. For example, the housingmay provide electricity and/or energy for the device. The housing maypower the device from an energy storage unit, energy generation unit,and/or energy conveyance unit of the housing. The housing may providecommunications between the device and/or an external device.

Controller

A controller may be provided at any level of the system describedherein. For example, one or more controller for a system, groups ofdevices, a single device, a module, a component of the device, and/or aportion of the component may be provided.

A system may comprise one or more controller. A controller may provideinstructions to one or more device, module of a device, component of adevice, and/or portion of a component. A controller may receive signalsthat may be detected from one or more sensors. A controller may receivea signal provided by a detection unit. A controller may comprise a localmemory or may access a remote memory. A memory may comprise tangiblecomputer readable media with code, instructions, language to perform oneor more steps as described elsewhere herein. A controller may be or usea processors.

A system wide controller may be provided external to one, two or moredevice and may provide instructions to or receive signals from the one,two or more devices. In some embodiments, the controller may communicatewith selected groups of devices. In some embodiments the controller maycommunicate with one or more devices in the same geographic location, orover different geographic locations. In some embodiments, a system widecontroller may be provided on a server or another network device. FIG.39 shows an example of a plurality of devices communicating with anexternal device over a network. In some instances, the external devicemay comprise a controller or be a controller communicating with theother devices. In some embodiments, a system wide controller may beprovided on a device, which may have a master-slave relationship withother devices.

In accordance with another embodiment of the invention, a device maycomprise one or more controller. The controller may provide instructionsto one or more module of the device, component of a device, and/orportion of a component. The device-level controller may receive signalsthat may be detected from one or more sensors, and/or a detection unit.

The controller may comprise a local memory or may access a remote memoryon the device. The memory may comprise tangible computer readable mediawith code, instructions, language to perform one or more steps asdescribed elsewhere herein. A device may have a local memory that maystore one or more protocols. In some embodiments, a controller may beprovided on a cloud computing infrastructure. The controller may bespread out across one or more hardware devices. The memory for thecontroller may be provided on one or more hardware devices. Theprotocols may be generated and/or stored on-board on the device.Alternatively, the protocols may be received from an external source,such as an external device or controller. The protocols may be stored ona cloud computing infrastructure, or a peer to peer infrastructure. Thememory may also store data collected from a detection unit of thedevice. The data may be stored for analysis of detected signals. Somesignal processing and/or data analysis may or may not occur at thedevice level. Alternatively, signal processing and/or data analysis mayoccur on an external device, such as a server. The signal processingand/or data analysis may occur using a cloud computing infrastructure.The signal processing and/or data analysis may occur at a differentlocation from where the device is located, or at the same geographiclocation.

The device-level controller may be provided within a device and mayprovide instructions to or receive signals from the one, two or moreracks, modules, components of a module, or portions of the components.In some embodiments, the controller may communicate with selected groupsof modules, components, or portions. In some instances, the device-levelcontroller may be provided within a module communicating with the othermodules. In some embodiments, a device-level controller may be providedon a module, which may have a master-slave relationship with othermodules. A modular controller may be insertable and/or removable from adevice.

A device level-controller may receive instructions from a system-widecontroller or a controller that provides instructions to one or moredevices. The instructions may be protocols which may be stored on alocal memory of the device. Alternatively, the instructions may beexecuted by the device in response to the received instructions withoutrequiring the instructions be stored on the device, or only having themtemporarily stored on the device. In some embodiments, the device mayonly store a recently received protocol. Alternatively, the device maystore multiple protocols and be able to refer to them at a later time.

The device may provide information related to detected signals from adetection unit to an external source. The external source receiving theinformation may or may not be the same as the source of the protocols.The device may provide raw information about the detected signals fromthe detection unit. Such information may include assay resultinformation. The device may provide some processing of the collectedsensor information. The device may or may not perform analysis of thecollected sensor information locally. The information sent to theexternal source may or may not include processed and/or analyzed data.

A device-level controller may instruct the device to perform as a pointof service device. A point of service device may perform one or moreaction at a location remote to another location. The device-levelcontroller may instruct the device to directly interface with a subjector environment. The device level controller may permit the device to beoperated by an operator of the device who may or may not be a healthcare professional. The device-level controller may instruct the deviceto directly receive a sample, where some additional analysis may occurremotely.

In accordance with additional embodiment of the invention, a module maycomprise one or more controller. The controller may provide instructionsto one or more components of the module, and/or portion of a component.The module-level controller may receive signals that may be detectedfrom one or more sensors, and/or a detection unit. In some examples,each module may have one or more controllers. Each module may have oneor multiple microcontrollers. Each module may have different operatingsystems that may control each module independently. The modules may becapable of operating independently of one another. One or more modulemay have one or more microcontrollers controlling different peripherals,detection systems, robots, movements, stations, fluid actuation, sampleactuation, or any other action within a module. In some instances, eachmodule may have built-in graphics capabilities for high performanceprocessing of images. In additional embodiments, each module may havetheir own controllers and/or processors that may permit parallelprocessing using a plurality of modules.

The controller may comprise a local memory or may access a remote memoryon the module. The memory may comprise tangible computer readable mediawith code, instructions, language to perform one or more steps asdescribed elsewhere herein. A module may have a local memory that maystore one or more protocols. The protocols may be generated and/orstored on-board on the module. Alternatively, the protocols may bereceived from an external source, such as an external module, device orcontroller. The memory may also store data collected from a detectionunit of the module. The data may be stored for analysis of detectedsignals. Some signal processing and/or data analysis may or may notoccur at the module level. Alternatively, signal processing and/or dataanalysis may occur on the device level, or at an external device, suchas a server. The signal processing and/or data analysis may occur at adifferent location from where the module is located, or at the samegeographic location.

The module-level controller may be provided within a module and mayprovide instructions to or receive signals from the one, two or morecomponents of the module, or portions of the components. In someembodiments, the controller may communicate with selected groups ofcomponents, or portions. In some instances, the module-level controllermay be provided within a component communicating with the othercomponents. In some embodiments, a module-level controller may beprovided on a component, which may have a master-slave relationship withother components. A modular controller may be insertable and/orremovable from a module.

A module-level controller may receive instructions from a device-widecontroller, system-wide controller or a controller that providesinstructions to one or more devices. The instructions may be protocolswhich may be stored on a local memory of the module. Alternatively, theinstructions may be executed by the module in response to the receivedinstructions without requiring the instructions be stored on the module,or only having them temporarily stored on the module. In someembodiments, the module may only store a recently received protocol.Alternatively, the module may store multiple protocols and be able torefer to them at a later time.

The module may provide information related to detected signals from adetection unit to the device, or an external source. The device orexternal source receiving the information may or may not be the same asthe source of the protocols. The module may provide raw informationabout the detected signals from the detection unit. Such information mayinclude assay result information. The module may provide some processingof the collected sensor information. The module may or may not performanalysis of the collected sensor information locally. The informationsent to the device or external source may or may not include processedand/or analyzed data.

A module-level controller may instruct the module to perform as a pointof service module. The module-level controller may instruct the moduleto directly interface with a subject or environment. The module levelcontroller may permit the module to be operated by an operator of thedevice who may or may not be a health care professional.

A controller may be provided at any level of the system as describedherein (e.g., high level system, groups of devices, device, rack,module, component, portion of component). The controller may or may nothave a memory at its level. Alternatively, it may access and/or use amemory at any other level. The controller may or may not communicatewith additional controllers at the same or different levels. Acontroller may or may not communicate with additional controllers atlevels immediately below or above them or a plurality of levels below orabove them. A controller may communicate to receive and/or provideinstructions/protocols. A controller may communicate to receive and/orprovide collected data or information based on the data.

User Interface

A device may have a display and/or user interface. In some situations,the user interface is provided to the subject with the aid of thedisplay, such as through a graphical user interface (GUI) that mayenable a subject to interact with device. Examples of displays and/oruser interfaces may include a touchscreen, video display, LCD screen,CRT screen, plasma screen, light sources (e.g., LEDs, OLEDs), IR LEDbased surfaces spanning around or across devices, modules or othercomponents, pixelsense based surface, infrared cameras or other capturetechnology based surfaces, projector, projected screen, holograms, keys,mouse, button, knobs, sliding mechanisms, joystick, audio components,voice activation, speakers, microphones, a camera (e.g., 2D, 3Dcameras), multiple cameras (e.g., may be useful for capturing gesturesand motions), glasses/contact lenses with screens built-in, videocapture, haptic interface, temperature sensor, body sensors, body massindex sensors, motion sensors, and/or pressure sensors. Any descriptionherein of a display and/or user interface may apply to any type ofdisplay and/or user interface. A display may provide information to anoperator of the device. A user interface may provide information and/orreceive information from the operator. In some embodiments, suchinformation may include visual information, audio information, sensoryinformation, thermal information, pressure information, motioninformation, or any other type of information. Sound, video, and colorcoded information (such as red LEDs indicating a module is in use) maybe used in providing feedback to users using a point of service systemor information system or interfacing with a system through touch orotherwise. In some embodiments, a user interface or other sensor of thedevice may be able to detect if someone is approaching the device, andwake up.

FIG. 56 illustrates a point of service device 5600 having a display5601. The display is configured to provide a graphical user interface(GUI) 5602 to a subject. The display 5601 may be a touch display, suchas a resistive-touch or capacitive-touch display. The device 5600 isconfigured to communicate with a remote device 5603, such as, forexample, a personal computer, Smart phone, tablet, or server. The device5600 has a central processing unit (CPU) 5604, memory 5605,communications module (or interface) 5606, and hard drive 5607. In someembodiments, the device 5600 includes a camera 5608 (or in some cases aplurality of cameras, such as for three-dimensional imaging) for imageand video capture. The device 5600 may include a sound recorder forcapturing sound. Images and/or videos may be provided to a subject withthe aid of the display 5601. In other embodiments, the camera 5608 maybe a motion-sensing input device (e.g., Microsoft® Kinect®).

One or more sensors may be incorporated into the device and/or userinterface. The sensors may be provided on the device housing, externalto the device housing, or within the device housing. Any of the sensortypes describing elsewhere herein may be incorporated. Some examples ofsensors may include optical sensors, temperature sensors, motionsensors, depth sensors, pressure sensors, electrical characteristicsensors, gyroscopes or acceleration sensors (e.g., accelerometer).

In an example, the device includes an accelerometer that detects whenthe device is not disposed on an ideal surface (e.g., horizontalsurface), such as when the device has tipped over. In another example,the accelerometer detects when the device is being moved. In suchcircumstances, the device may shutdown to prevent damage to variouscomponents of the device. In some cases, prior to shutting down, thedevice takes a picture of a predetermined area on or around the devicewith the aid of a camera on the device (see FIG. 56).

The user interface and/or sensors may be provided on a housing of thedevice. They may be integrated into the housing of a device. In someembodiments, the user interface may form an outer layer of the housingof the device. The user interface may be visible when viewing thedevice. The user interface may be selectively viewable when operatingthe device.

The user interface may display information relating to the operation ofthe device and/or data collected from the device. The user interface maydisplay information relating to a protocol that may run on the device.The user interface may include information relating to a protocolprovided from a source external to the device, or provided from thedevice. The user interface may display information relating to a subjectand/or health care access for the subject. For example, the userinterface may display information relating to the subject identity andmedical insurance for the subject. The user interface may displayinformation relating to scheduling and/or processing operation of thedevice.

The user interface may be capable of receiving one or more input from auser of the device. For example, the user interface may be capable ofreceiving instructions about one or more assay or procedure to beperformed by the device. The user interface may receive instructionsfrom a user about one or more sample processing step to occur within thedevice. The user interface may receive instructions about one or moreanalyte to be tested for.

The user interface may be capable of receiving information relating tothe identity of the subject. The subject identity information may beentered by the subject or another operator of the device or imaged orotherwise captured by the user interface itself. Such identification mayinclude biometric information, issued identification cards, or otheruniquely identifiable biological or identifying features, materials, ordata. The user interface may include one or more sensors that may assistwith receiving identifying information about the subject. The userinterface may have one or more question or instructions pertaining tothe subject's identity, to which the subject may respond.

In some situations, the user interface is configured to display aquestionnaire to a subject, the questionnaire including questions aboutthe subject's dietary consumption, exercise, health condition and/ormental condition (see above). The questionnaire may be a guidedquestionnaire, having a plurality of questions of or related to thesubject's dietary consumption, exercise, health condition and/or mentalcondition. The questionnaire may be presented to the subject with theaid of a user interface, such as graphical user interface (GUI), on thedisplay of the device.

The use interface may be capable of receiving additional informationrelating to the subject's condition, habits, lifestyle, diet, exercise,sleep patterns, or any other information. The additional information maybe entered directly by the subject or another operator of the device.The subject may be prompted by one or more questions or instructionsfrom the user interface and may enter information in response. Thequestions or instructions may relate to qualitative aspects of thesubject's life (e.g., how the patient is feeling). In some embodiments,the information provided by the subject are not quantitative. In someinstances, the subject may also provide quantitative information.Information provided by the subject may or may not pertain to one ormore analyte level within a sample from the subject. The survey may alsocollect information relating to therapy and/or medications undergone orcurrently taken by the subject. The user interface may prompt thesubject using a survey or similar technique. The survey may includegraphics, images, video, audio, or other media features. The survey mayor may not have a fixed set of questions and/or instructions. The survey(e.g., the sequence and/or content of the questions) may dynamicallychange depending on the subject's answers.

Identifying information about the subject and/or additional informationrelating to the subject may be stored in the device and/or transmittedto an external device or cloud computing infrastructure. Suchinformation may be useful in analyzing data relating to a samplecollected from the subject. Such information may also be useful fordetermining whether to proceed with sample processing.

The user interface and/or sensors may be capable of collectinginformation relating to the subject or the environment. For example, thedevice may collect information through a screen, thermal sensor, opticalsensor, motion sensor, depth sensor, pressure sensor, electricalcharacteristic sensor, acceleration sensor, any other type of sensordescribed herein or known in the art. In one example, the optical sensormay be a multi-aperture camera capable of collecting a plurality ofimages and calculating a depth therefrom. An optical sensor may be anytype of camera or imaging device as described elsewhere herein. Theoptical sensor may capture one or more static images of the subjectand/or video images of the subject.

The device may collect an image of the subject. The image may be a 2Dimage of the subject. The device may collect a plurality of images ofthe subject that may be used to determine a 3D representation of thesubject. The device may collect a one-time image of the subject. Thedevice may collect images of the subject over time. The device maycollect images with any frequency. In some embodiments, the device maycontinually collect images in real-time. The device may collect a videoof the subject. The device may collect images relating to any portion ofthe subject including but not limited to the subject's eye or retina,the subject's face, the subject's hand, the subject's fingertip, thesubject's torso, and/or the subject's overall body. The images collectedof the subject may be useful for identifying the subject and/or fordiagnosis, treatment, monitoring, or prevention of a disease for thesubject. In some instances, images may be useful for determining thesubject's height, circumference, weight, or body mass index. The devicemay also capture the image of a subject's identification card, insurancecard, or any other object associated with the subject.

The device may also collect audio information of the subject. Such audioinformation may include the subject's voice or the sound of one or morebiological process of the subject. For example, the audio informationmay include the sound of the subject's heartbeat.

The device may collect biometric information about a subject. Forexample, the device may collect information about the subject's bodytemperature. In another example, the device can collect informationabout the subject's pulse rate. In some instances, the device may scan aportion of the subject, such as the subject's retina, fingerprint orhandprint. The device may determine the subject's weight. The device mayalso collect a sample from the subject and sequence the subject's DNA ora portion thereof. The device may also collect a sample from the subjectand conduct a proteomic analysis thereon. Such information may be usedin the operation of the device. Such information may relate to thediagnosis or the identity of the subject. In some embodiments, thedevice may collect information about the operator of the device who mayor may not be different from the subject. Such information can be usefulfor verifying the identity of the operator of the device.

In some instances, such information collected by the device may be usedto identify the subject. The subject's identity may be verified forinsurance or treatment purposes. The subject identify may be tied to thesubject's medical records. In some instances, the data collected by thedevice from the subject and/or sample may be linked to the subject'srecords. The subject identity may also be tied into the subject's healthinsurance (or other payer) records.

Power Source

A device may have a power source or be connected to a power source. Insome embodiments, the power source may be provided external to thedevice. For example, the power may be provided from a grid/utility. Thepower may be provided from an external energy storage system or bank.The power may be provided by an external energy generation system. Insome embodiments, the device may include a plug or other connectorcapable of electrically connecting the device to the external powersource. In another example, the device may use a body's naturalelectrical impulses to power the device. For example, the device maycontact a subject, be worn by the subject, and/or be ingested by thesubject, who may or may not provide some power to the device. In someembodiments, the device may include one or more piezoelectric componentthat may be movable, and capable of providing power to the device. Forexample, the device may have a patch configuration configured to beplaced on a subject, so that when the subject moves and/or the patch isflexed, power is generated and provided to the device.

A device may optionally have an internal power source. For example, alocal energy storage may be provided on the device. In one embodiment,the local energy storage may be one or more battery or ultracapacitor.Any battery chemistry known or later developed in the art may be used asa power source. A battery may be a primary or secondary (rechargeable)battery. Examples of batteries may include, but are not limited to,zinc-carbon, zinc-chloride, alkaline, oxy-nickel hydroxide, lithium,mercury oxide, zinc-air, silver oxide, NiCd, lead acid, NiMH, NiZn, orlithium ion. The internal power source may be stand alone or may becoupled with an external power source. In some embodiments, a device mayinclude an energy generator. The energy generator may be provided on itsown or may be coupled with an external and/or internal power source. Theenergy generator may be a traditional electricity generator as known inthe art. In some embodiments, the energy generator may use a renewableenergy source including, but not limited to, photovoltaics, solarthermal energy, wind energy, hydraulic energy, or geothermal energy. Insome embodiments, the power may be generated through nuclear energy orthrough nuclear fusion.

Each device may be connected to or have a power source. Each module maybe connected to or have its own local power source. In some instances,modules may be connected to a power source of the device. In someinstances, each module may have its own local power source and may becapable of operating independently of other modules and/or devices. Insome instances, the modules may be able to share resources. For example,if a power source in one of the modules is damaged or impaired, themodule may be able to access the power source of another module or ofthe device. In another example, if a particular module is consuming alarger amount of power, the module may be able to tap into the powersource of another module or of the device.

Optionally, device components may have a power source. Any discussionherein relating to power sources of modules and/or devices may alsorelate to power sources at other levels, such as systems, groups ofdevices, racks, device components, or portions of device components.

Communication Unit

A device may have a communication unit. The device may be capable ofcommunication with an external device using the communication unit. Insome instances, the external device may be one or more fellow devices.The external device may be a cloud computing infrastructure, part of acloud computing infrastructure, or may interact with a cloud computinginfrastructure. In some instances, the external device that the devicemay communicate with may be a server or other device as describedelsewhere herein.

The communication unit may permit wireless communication between thedevice and the external device. Alternatively, the communication unitmay provide wired communication between the device and the externaldevice. The communication unit may be capable of transmitting and/orreceiving information wirelessly from an external device. Thecommunication unit may permit one way and/or two-way communicationbetween the device and one or more external device. In some embodiments,the communication unit may transmit information collected or determinedby the device to an external device. In some embodiments, thecommunication unit may be receiving a protocol or one or moreinstructions from the external device. The device may be able tocommunicate with selected external devices, or may be able tocommunicate freely with a wide variety of external devices.

In some embodiments, the communication unit may permit the device tocommunicate over a network, such as a local area network (LAN) or widearea network (WAN) such as the Internet. In some embodiments, the devicemay communicate via a telecommunications network, such as a cellular orsatellite network.

Some examples of technologies that may be used by a communication unitmay include Bluetooth or RTM technology. Alternatively, variouscommunication methods may be used, such as a dial-up wired connectionwith a modem, a direct link such as TI, ISDN, or cable line. In someembodiments, a wireless connection may be using exemplary wirelessnetworks such as cellular, satellite, or pager networks, GPRS, or alocal data transport system such as Ethernet or token ring over a LAN.In some embodiments, the communication unit may contain a wirelessinfrared communication component for sending and receiving information.

In some embodiments, the information may be encrypted before it istransmitted over a network, such as a wireless network. In someembodiments, the encryption may be hardware-based encryption. In someinstances, the information may be encrypted on the hardware. Any or allinformation, which may include user data, subject data, test results,identifier information, diagnostic information, or any other type ofinformation, may be encrypted based on hardware based and/or softwarebased encryption. Encryption may also optionally be based onsubject-specific information. For example, a subject may have a samplebeing processed by the device, and the subject's password may be used toencrypt the data relating to the subject's sample. By encrypting thesubject's data with subject-specific information, only the subject maybe able to retrieve that data. For example, the decryption may onlyoccur if the subject enters a password on a website. In another example,information transmitted by the device may be encrypted by informationspecific to the operator of the device at that time, and may only beretrieved if the operator enters the operator's password or provide theoperator specific-information.

Each device may have a communication unit. Each module may have its ownlocal communication unit. In some instances, modules may share acommunication unit with the device. In some instances, each module mayhave its own local communication unit and may be capable ofcommunicating independently of other modules and/or devices. The modulemay use its communication unit to communicate with an external device,with the device, or with other modules. In some instances, the modulesmay be able to share resources. For example, if a communication unit inone of the modules is damaged or impaired, the module may be able toaccess the communication unit of another module or of the device. Insome instances, devices, racks, modules, components or portions ofdevice components may be able to share one or more routers. The variouslevels and/or components in the hierarchy may be able to communicatewith one another.

Optionally, device components may have a communication unit. Anydiscussion herein relating to communication units of modules and/ordevices may also relate to communication units at other levels, such assystems, groups of devices, racks, device components, or portions ofdevice components.

Device, Module and Component Identifier

A device may have a device identifier. A device identifier may identifythe device. In some embodiments, the device identifier may be unique perdevice. In other embodiments, the device identifier may identify a typeof device, or modules/components provided within the device. The deviceidentifier may indicate functions that the device is capable ofperforming. The device identifier may or may not be unique in suchsituations.

The device identifier may be a physical object formed on the device. Forexample, the device identifier may be read by an optical scanner, or animaging device, such as a camera. The device identifier may be read byone or more types of sensors described elsewhere herein. In one example,the device identifier may be a barcode. A barcode may be a 1D or 2Dbarcode. In some embodiments, the device identifier may emit one or moresignal that may identify the device. For example, the device identifiermay provide an infrared, thermal, ultrasonic, optical, audio,electrical, chemical, biological, or other signal that may indicate thedevice identity. The device identifier may use a radiofrequencyidentification (RFID) tag.

The device identifier may be stored in a memory of the device. In oneexample, the device identifier may be a computer readable medium. Thedevice identifier may be communicated wirelessly or via a wiredconnection.

The device identifier may be static or changeable. The device identifiermay change as one or more module provided for the device may change. Thedevice identifier may change based on available components of thedevice. The device identifier may change when instructed by an operatorof the device.

The device identifier may be provided to permit the device to beintegrated within a systemwide communication. For example, an externaldevice may communicate with a plurality of devices. The external devicemay distinguish a diagnostic device from another diagnostic device viathe device identifier. The external device may provide specializedinstructions to a diagnostic device based on its identifier. Theexternal device may include a memory or may communicate with a memorythat may keep track of information about the various devices. The deviceidentifier of a device may be linked in memory with the informationcollected from the device or associated with the device.

In some embodiments, an identifier may be provided on a module or atcomponent level to uniquely identify each component in a device at thesystem level. For example, various modules may have module identifiers.The module identifier may or may not be unique per module. The moduleidentifier may have one or more characteristics of a device identifier.

The module identifier may permit a device or system (e.g., externaldevice, server) to identify the modules that are provided therein. Forexample, the module identifier may identify the type of module, and maypermit the device to automatically detect the components and capabilityprovided by the module. In some instances, the module identifier mayuniquely identify the module, and the device may be able to trackspecific information associated with the particular module. For example,the device may be able to track the age of the module and estimate whencertain components may need to be renewed or replaced. The module maycommunicate with a processor of the device which it is a part of.

Alternatively, the module may communicate with a processor of anexternal device. The module identifier may provide the same informationon a system-wide level. In some embodiments, the system, rather than thedevice, may track the information associated with the module identifier.

The module identifier may be communicated to the device or system whenit is connected to the device or interfaced with a device. For instance,the module identifier may be communicated to the device or system afterthe module has been mounted on a support structure. Alternatively, themodule identifier may be communicated remotely when the module is notyet connected to the device.

An identifier may be provided at any other level described herein (e.g.,external device, groups of devices, racks, components of a device,portions of a component). Any characteristics of identifiers providedherein may also apply to such identifiers.

Systems

FIG. 39 provides an illustration of a diagnostic system in accordancewith an embodiment of the invention. One, two or more devices 3900 a,3900 b may communicate with an external device 3910 over a network 3920.The devices may be diagnostic devices. The devices may have any featuresor characteristics as described elsewhere herein. In some examples, thedevices may be a benchtop device, handheld device, patch, and/or pill.The devices may be configured to accept a sample and perform one or moreof a sample preparation step, assay step, or detection step. The devicesmay comprise one or more modules as described elsewhere herein.

In some embodiments, a patch or pill is configured to be operativelycoupled (or linked) to a mobile device, such as a network device, thatis configured to communicate with another device and/or a network (e.g.,intranet or the Internet). In some situations, a patch is configured tocommunicate with a pill circulating through the body of a subject, ordisposed in the body of the subject, such as in a tissue of the subject.In other situations, a pill is a particle having a size on the order ofnanometers, micrometers or larger. In an example, a pill is ananoparticle. The patch and/or pill may include onboard electronics topermit the patch and/or pill to communicate with another device.

A system may include any number of devices 3900 a, 3900 b. For example,the system may include one or more, two or more, three or more, four ormore, five or more, ten or more, twenty or more, fifty or more, onehundred or more, five hundred or more, one thousand or more, fivethousand or more, ten thousand or more, one hundred thousand or more, orone million or more devices.

The devices may or may not be associated into groups of devices. Adevice may be associated with one, two, three, ten or any number ofgroups. A device may be part of groups, sub-groups, sub-sub-groups withno limitations of sub-grouping in the system. In some embodiments,groups of devices may include devices at a particular geographiclocation. For example, groups of devices may refer to devices within thesame room or within the same building. A group of devices may includedevices within the same retailer location, laboratory, clinic, healthcare facility, or any other location. Groups of devices may refer todevices within the same town or city. Groups of devices may includedevices within a particular radius. In some instances, groups of devicesmay include devices using the same communication port. For example,groups of devices may include devices using the same router, Internethub, telecommunications tower, satellite, or other communication port.

Alternatively, groups of devices may include devices associated with thesame entity or division of an entity. For example, a group of devicesmay be associated with a laboratory, health care provider, medicalfacility, retailer, company, or other entity.

Any description herein on a system-wide level may refer to an overallglobal system that may include or communicate with any device.Alternatively, any description herein of a system may also refer to agroup of devices.

A network 3920 may be provided, as described elsewhere herein. Forexample, the network may include a local area network (LAN) or wide areanetwork (WAN) such as the Internet. In some embodiments, the device maycommunicate via a telecommunications network, such as a cellular orsatellite network.

A device may communicate with the network using a wireless technology,such as Bluetooth or RTM technology. Alternatively, variouscommunication methods may be used, such as a dial-up wired connectionwith a modem, a direct link such as TI, ISDN, or cable line. In someembodiments, a wireless connection may be using exemplary wirelessnetworks such as cellular, wimax, wifi, satellite, or pager networks,GPRS, or a local data transport system such as Ethernet or token ringover a LAN. In some embodiments, the device may communicate wirelesslyusing infrared communication components.

An external device 3910 may be provided in accordance with an embodimentof the invention. The external device may be any networked devicedescribed elsewhere herein or known in the art. For example, theexternal device may be a server, personal computer, laptop computer,tablet, mobile device, cell phone, satellite phone, smart phone (e.g.,iPhone, Android, Blackberry, Palm, Symbian, Windows), personal digitalassistant (PDA), pager or any other device. In some instances, theexternal device may be another diagnostic device. A master-slaverelationship, peer-to-peer or a distributed relationship, may beprovided between the diagnostic devices.

The external device may have a processor and memory. The external devicemay access a local memory or communicate with a memory. The memory mayinclude one or more databases.

Any description of the external device may also apply to any cloudcomputing infrastructure. An external device may refer to one or moredevices that may include processors and/or memory. The one or moredevices may or may not be in communication with one another.

In some embodiments, the external device may function as a controller ormay comprise a controller, and perform one or more functions of thecontroller as described elsewhere herein. The external device mayfunction as a system-wide controller, may control a group of devices, ormay control an individual device.

In one example, an external device may have data stored in memory. Suchdata may include analyte threshold data. Such data may include curves orother information that may be useful for performing analysis and/orcalibration. The external device may also receive and/or store datareceived from a sample processing device. Such data may include datarelated to one or more signals detected by the sample processing device.In some embodiments, one or more diagnostics and/or calibrations may beperformed on the sample processing device. Such diagnostics and/orcalibrations may use and/or access curves or other data stored on-boardthe device or at an external device, such as a server.

FIG. 1 shows an example of a device 100 in communication with acontroller 110 in accordance with an embodiment of the invention.

The device may have any of the characteristics, structure, orfunctionality as described elsewhere herein. For example, the device 100may comprise one or more support structure 120. In some embodiments, thesupport structure may be a rack, or any other support as describedelsewhere herein. In some instances, the device may include a singlesupport structure. Alternatively, the device may include a plurality ofsupport structures. A plurality of support structure may or may not beconnected to one another.

The device 100 may comprise one or more module 130. In some instances, asupport structure 120 may comprise one or more module. In one example,the module may have a blade format that may be mounted on a rack supportstructure. Any number of modules may be provided per device or supportstructure. Different support structure may have different numbers ortypes of modules.

The device 100 may comprise one or more component 140. In someinstances, a module 130 may comprise one or more component of themodule. A rack 120 may comprise one or more component of a module. Anynumber of components may be provided per device, rack, or module.Different modules may have different numbers or types of components.

In some examples, the devices may be a benchtop device, a handhelddevice, a wearable device, an ingestible device, an implantable device,a patch, and/or a pill. The device may be portable. The device may beplaced on top of a surface, such as a counter, table, floor or any othersurface. The device may be mountable or attachable to a wall, ceiling,ground and/or any other structure. The device may be worn directly bythe subject, or may be incorporated into the subject's clothing.

The device may be self-contained. For example, the device may comprise alocal memory. The local memory may be provided to the overall device, ormay be provided to one or more module, or may be distributed over one ormore module. The local memory may be contained within a housing of thedevice. A local memory may be provided on a support of a module orwithin a housing of a module. Alternatively, the local memory of thedevice may be provided external to a module while within the devicehousing. The local memory of the device may or may not be supported by asupport structure of the device. The local memory may be providedexternal to the support structure of the device, or may be integratedwithin the support structure of the device.

One or more protocols may be stored in a local memory. One or moreprotocols may be delivered to the local memory. The local memory mayinclude a database of information for on board analysis of detectedsignals. Alternatively, the local memory may store the informationrelated to the detected signals that may be provided to an externaldevice for remote analysis. The local memory may include some signalprocessing of the detected signals, but may be transmitted to theexternal device for analysis. The external device may or may not be thesame device the controller.

The local memory may be capable of storing non-transitory computerreadable media, which may include code, logic, or instructions capableof performing steps described herein.

The device may comprise a local processor. The processor may be capableof receiving instructions and providing signals to execute theinstructions. The processor may be a central processing unit (CPU) thatmay carry out instructions of tangible computer readable media. In someembodiments, the processor may include one or more microprocessors. Theprocessor may be capable of communicating with one or more component ofthe device, and effecting the operation of the device.

The processor may be provided to the overall device, or may be providedto one or more module, or may be distributed over one or more module.The processor may be contained within a housing of the device. Aprocessor may be provided on a support of a module or within a housingof a module. Alternatively, the processor of the device may be providedexternal to a module while within the device housing. The processor ofthe device may or may not be supported by a support structure of thedevice. The processor may be provided external to the support structureof the device, or may be integrated within the support structure of thedevice.

A controller 110 may be in communication with the device 100. In someembodiments, the controller may be a system-wide controller. Thecontroller may communicate with any device. The controller may beselectively in communication with a group of devices. For example, thesystem may comprise, one, two or more controller, wherein a controllermay be devoted to a group of devices. The controller may be capable ofindividually communicating with each device. In some instances, thecontroller may communicate with groups of devices, withoutdifferentiating between the devices within the group. The controller maycommunicate with any combination of devices or groups of devices.

A controller may be provided external to the device. The controller maybe an external device in communication with the device. As describedelsewhere herein, an external device may be any sort of network device.For example the controller may be a server, a mobile device, or anotherdiagnostic device which may have a master-slave relationship with thedevice.

In alternate embodiments, the controller may be provided locally to thedevice. In such situations, the device may be entirely self-containedwithout requiring external communication.

The controller may comprise a memory or may communicate with a memory.One or more protocols may be stored on the controller memory. Theseprotocols may be stored external to the device. The protocols may bestored in a memory and/or cloud computing infrastructure. The protocolsmay be updated on the controller side without having to modify thedevice. The controller memory may include a database of informationrelating to devices, samples, subjects, and/or information collectedfrom the devices. The information collected from the devices may includeraw data of detected signals within the device. The informationcollected from the devices may include some signal processing of thedetected signals. Alternatively, the information collected from thedevices may include analysis that may have been performed on board thedevice.

The controller memory may be capable of storing non-transitory computerreadable media, which may include code, logic, or instructions capableof performing steps described herein.

The controller may comprise a processor. The processor may be capable ofreceiving instructions and providing signals to execute theinstructions. The processor may be a central processing unit (CPU) thatmay carry out instructions of tangible computer readable media. In someembodiments, the processor may include one or more microprocessors. Theprocessor of the controller may be capable of analyzing data receivedfrom the devices. The processor of the controller may also be capable ofselecting one or more protocol to provide to the device.

In some embodiments, the controller may be provided on a single externaldevice. The single external device may be capable of providing protocolsto the diagnostic device and/or receiving information collected from thediagnostic device. In some instances, the controller may be providedover a plurality of devices. In one example, a single external device ormultiple external devices may be capable of providing protocols to thediagnostic device. A single external device or multiple external devicesmay be capable of receiving information collected from the diagnosticdevice. A single external device or multiple external devices may becapable of analyzing the information collected from the diagnosticdevice.

Alternatively, the system may use cloud computing. One or more functionsof the controller may be provided by a computer network, rather thanbeing limited to a single external device. In some embodiments, anetwork or plurality of external devices may communicate with thediagnostic device and provide instructions to, or receive informationfrom the diagnostic device. Multiple processors and storage devices maybe used to perform the functions of the controller. The controller maybe provided in an environment enabling convenient, on-demand networkaccess to a shared pool of configurable computing resources (e.g.,networks, servers, storage, applications, and services) that can berapidly provisioned and released with minimal management effort orservice provider interaction.

Communication may be provided between a diagnostic device and acontroller. The communication may be one way communication. For example,the controller may push down a protocol to the device. In anotherexample, the device may initiate a request for a protocol from thecontroller. Or the device may only provide information to the controllerwithout requiring a protocol from the controller.

Preferably, two-way communication may be provided between the diagnosticdevice and the controller. For example, a protocol may be provided froma source external to the device. The protocol may or may not be based oninformation provided by the device. For example, the protocol may or maynot be based on an input provided to the device, which may somehowdetermine the information provided by the device to the controller. Theinput may be manually determined by an operator of the device. Forexample, the operator may specify one or more tests that the operatorwishes the device to perform. In some instances, the input may bedetermined automatically. For example, the tests to run may bedetermined automatically based on a characteristic of the sample, whichmodules are available or used, past records relating to a subject, aschedule of anticipated tests, or any other information.

In some embodiments, the device may request specific protocols from thecontroller. In some other embodiments, the device may provideinformation to the controller, and the controller may select one or moreprotocols to provide to the device based on that information.

The device may provide information collected at the device based on oneor more detected signals from one or more sensors. The sensedinformation may be provided to the controller. The sensed informationmay or may not be collected during the operation of a protocol. In someembodiments, the controller may provide an additional protocol based onthe information collected during the first protocol. The first protocolmay be completed before the additional protocol is initiated, or theadditional protocol may be initiated before the first protocol iscompleted, based on the information collected.

A feedback system may be provided wherein a protocol may be provided oraltered based on information collected during a protocol or after thecompletion of a protocol. One or more protocol may run in parallel, insequence, or in any combination thereof. A device may perform aniterative process, which may use instructions, actions performed basedon the instructions, data collected from the actions performed, whichmay optionally affect subsequent instructions, and so forth. A protocolmay cause the device to perform one or more action, including but notlimited to, a sample collection step, sample preparation step, assaystep, and/or detection step.

Within a system, a device may be capable of communicating with one ormore entity. For example, the device may communicate with a lab benefitsmanager, who may collect information from the device. The lab benefitsmanager may analyze the information collected from the device. Thedevice may communicate with a protocol provider, who may provide one ormore instructions to the device. The protocol provider and lab benefitsmanager may be the same entity, or may be different entities. The devicemay optionally communicate with a payer, such as an insurance company.The device may optionally communicate with a health care provider. Thedevice may communicate directly with one or more of these entities, ormay communicate with them indirectly through another party. In oneexample, the device may communicate with a lab benefits manager, who maycommunicate with a payer and health care provider.

In some embodiments, the device may enable a subject to communicate witha health care provider. In one example, the device may permit one ormore image of a subject to be taken by the device, and provided to thesubject's physician. The subject may or may not view the physician onthe device. The image of the subject may be used to identification ordiagnostic purposes. Other information relating to the subject'sidentification may be used, as described elsewhere herein. The subjectmay communicate with the physician in real-time. Alternatively, thesubject may view a recording provided by the physician. The subject mayadvantageously be communicating with the subject's own physician whichmay provide additional comfort and/or sense of personal interaction forthe subject. Alternatively, the subject may communicate with otherhealth care providers, such as specialists.

In some embodiments, diagnostic devices within a system may shareresources. For example devices within a system may be communicating withone another. The devices may be directly linked to one another, or maycommunicate over a network. The devices may be directly linked to ashared resource or may communicate over a network with the sharedresource. An example of a shared resource may be a printer. For example,a plurality of devices may be in communication with a single printer.Another example of a shared resource may be a router.

A plurality of devices may share additional peripherals. For example, aplurality of devices within a system may communicate with a peripheralthat may capture one or more physiological parameter of a subject. Forexample, the devices may communicate with a blood pressure measuringdevice, a scale, a pulse rate measuring device, and ultrasound imagecapturing device, or any other peripheral device. In some instances, aplurality of devices and/or systems may communicate with a computer,mobile device, tablet, or any other device that may be useful forinterfacing with a subject. Such external devices may be useful forcollecting information from the subject via a survey. In someembodiments, one or more controller of a system may determine whichdevice may be using which peripheral at any given moment. In someembodiments, a peripheral device may communicate with a sampleprocessing device a wireless connection (e.g. Bluetooth).

The system may be capable of dynamic resource allocation. In someembodiments, the dynamic resource allocation may be system-wide orwithin a group of devices. For example, a plurality of devices may beconnected to a plurality of shared resources. In one example, devices Aand B may be connected to printer X, and devices C and D may beconnected to printer Y. If a problem occurs with printer X, devices Aand B may be able to use printer Y. Devices A and B may be able tocommunicate directly with printer Y. Alternatively, devices A and B maynot be able to communicate directly with printer Y, but may be able tocommunicate with printer Y through devices C and D. The same may go forrouters, or other sharable resources.

Methods

Methods for Processing Samples

In some embodiments, a single device, such as a module or a systemhaving one or more modules, is configured to perform one or moreroutines selected from the group consisting of sample preparation,sample assaying and sample detection. Sample preparation may includephysical processing and chemical processing. The single device in somecases is a single module. In other cases, the single device is a systemhaving a plurality of modules, as described above.

FIG. 40 shows an example of one or more step that may be performed in amethod. The method may or may not be performed by a single device.

The method may include the step of sample collection 4000, samplepreparation 4010, sample assay 4020, detection 4030, and/or output 4040.Any of these steps may be optional. Furthermore, these steps may occurin any order. One or more of the steps may be repeated one or moretimes.

In one example, after a sample is collected, it may undergo one or moresample preparation step. Alternatively, after the sample is collected,it may directly go to a sample assay step. In another example, adetection step may occur directly after the sample is collected. In oneexample, the detection step may include taking an image of the sample.The image may be a digital image and/or video.

In another example, after a sample has undergone one or more samplepreparation step, it may go to a sample assay step. Alternatively, itmay go directly to a detection step.

After a sample has undergone one or more assay step, the sample mayproceed to a detection step. Alternatively, the sample may return to oneor more sample preparation step.

After a sample has undergone a detection step, it may be output.Outputting may include displaying and/or transmitting data collectedduring the detection step. Following detection, the sample may undergoone or more sample preparation step or sample assay step. In someinstances, following detection, additional sample may be collected.

After a sample has been displayed and/or transmitted, additional samplepreparation steps, sample assay steps, and/or detection steps may beperformed. In some instances, protocols may be sent to a device inresponse to transmitted data, which may effect additional steps. In someinstances, protocols may be generated on-board in response to detectedsignals. Analysis may occur on-board the device or may occur remotelybased on transmitted data.

A single device may be capable of performing one or more sampleprocessing steps. In some embodiments, the term “processing” encompassesone or more of preparing the sample, assaying the sample, and detectingthe sample to generate data for subsequent analysis off-board (i.e., offthe device) or on-board (i.e., on the device). A sample processing stepmay include a sample preparation procedure and/or assay, including anyof those described elsewhere herein. Sample processing may include oneor more chemical reactions and/or physical processing steps describedherein. Sample processing may include the assessment of histology,morphology, kinematics, dynamics, and/or state of a sample, which mayinclude such assessment for cells or other assessment described herein.In an embodiment, a single device is configured to one or more samplepreparation procedures selected from the group consisting of weighing orvolume measurement of the sample, centrifugation, sample processing,separation (e.g., magnetic separation), other processing with magneticbeads and/or nanoparticles, reagent processing, chemical separation,physical separation, chemical separation, incubation, anticoagulation,coagulation, removal of parts of sample (e.g., physical removal ofplasma, cells, lysate), dispersion/dissolution of solid matter,concentration of selected cells, dilution, heating, cooling, mixing,addition of reagent(s), removal of interfering factors, preparation of acell smear, pulverization, grinding, activation, ultrasonication, microcolumn processing, and/or any other type of sample preparation stepknown in the art, including but not limited to those listed in FIG. 57.In an example, a single module is configured to perform multiple samplepreparation procedures. In another example, a single system, such as thesystem 700, is configured to perform multiple sample preparationprocedures. In another embodiment, a single device is configured toperform 1 or more, or 2 or more, or 3 or more, or 4 or more, or 5 ormore, or 10 or more assays selected from the group consisting ofimmunoassay, nucleic acid assay, receptor-based assay, cytometric assay,colorimetric assay, enzymatic assay, electrophoretic assay,electrochemical assay, spectroscopic assay, chromatographic assay,microscopic assay, topographic assay, calorimetric assay, turbidimetricassay, agglutination assay, radioisotope assay, viscometric assay,coagulation assay, clotting time assay, protein synthesis assay,histological assay, culture assay, osmolarity assay, and/or other typesof assays or combinations thereof. In some situations, a single deviceis configured to perform multiple types of assays, at least one of whichis cytometry or agglutination. In other situations, a single device isconfigured to perform multiple types of assays, including cytometry andagglutination. In an example, the system 700 is configured to performcytometry with the aid of the cytometry station 707. A single device maybe configured to perform any number of assays, including the numbersdescribed elsewhere herein, in areas relating to Chemistry—RoutineChemistry, Hematology (includes cell-based assays, coagulation andandrology), Microbiology—Bacteriology (includes “Molecular Biology”),Chemistry—Endocrinology, Microbiology—Virology, DiagnosticImmunology—General Immunology, Chemistry—Urinalysis,Immunohematology—ABO Group & Rh type, Diagnostic Immunology—SyphilisSerology, Chemistry—Toxicology, Immunohematology—Antibody Detection(transfusion), Immunohematology—Antibody Detection (non-transfusion),Histocompatibility, Microbiology—Mycobacteriology,Microbiology—Mycology, Microbiology—Parasitology,Immunohematology—Antibody Identification, Immunohematology—CompatibilityTesting, Pathology—Histopathology, Pathology—Oral Pathology,Pathology—Cytology, Radiobioassay, or Clinical Cytogenetics. The singledevice may be configured for the measurement of one or more or, two ormore of, three or more of, or any number of (including those describedelsewhere herein): proteins, nucleic acids (DNA, RNA, hybrids thereof,microRNA, RNAi, EGS, Antisense), metabolites, gasses, ions, particles(which may include crystals), small molecules and metabolites thereof,elements, toxins, enzymes, lipids, carbohydrates, prion, formed elements(e.g., cellular entities (e.g., whole cell, cell debris, cell surfacemarkers)). A single device may be capable of performing various types ofmeasurements, including but not limited to imaging,spectrometry/spectroscopy, electrophoresis, chromatography,sedimentation, centrifugation, or any others mentioned in FIG. 58.

In some situations, the histology of a sample encompasses staticinformation of the sample as well as temporal change of the sample. Inan example, the sample as collected contains cells that multiply (ordivide) or metastasize after the sample is collected.

In another embodiment, a single device is configured to perform one ormore types of sample detection routines, such as those describedelsewhere herein.

In some embodiments, multi-use or multi-purpose devices are configuredto prepare and process a sample. Such devices may include 1 or more, or2 or more, or 3 or more, or 4 or more, or 5 or more, or 6 or more, or 7or more, or 8 or more, or 9 or more, or 10 or more, or 20 or more, or 30or more, or 40 or more, or 50 or more, or 100 or more modules, either aspart of a single system or a plurality of systems in communication withone another. The modules may be in fluid communication with one another.Alternatively, the modules may be fluidically isolated or hydraulicallyindependent from one another. In such a case, a sample transfer devicemay enable transferring a sample to and from a module. Such devices mayaccept 1 or more, or 2 or more, or 3 or more, or 4 or more, or 5 ormore, or 6 or more, or 7 or more, or 8 or more, or 9 or more, or 10 ormore, or 20 or more, or 30 or more, or 40 or more, or 50 or more, or 100or more samples. In an embodiment, devices accept samples in a batchfashion (e.g., 5 samples provided to a device at once). In anotherembodiment, devices accept samples in a continuous fashion. In someembodiments, fluidically isolated or hydraulically independent modulesare hydraulically isolated from one another.

In an embodiment, samples are processed in parallel. In anotherembodiment, samples are processed sequentially (or one after another).Devices provided herein may prepare and analyze the same sample or aplurality of different samples. In an example, devices provided hereinprocess the same blood, urine and/or tissue sample. In another example,devices provided herein process different blood, urine and/or tissuesamples.

In some embodiments, devices for processing samples accept samples ofvolumes of at least about 1 nanoliter (nL), or 10 nL, or 100 nL, or 1microliter (μL), or 10 μL, or 100 μL, or 1 milliliter (mL), or 10 mL, or100 mL, or 1 liter (L), or 2 L, or 3 L, or 4 L, or 5 L, or 6 L, or 7 L,or 8 L, or 9 L, or 10 L, or 100 L, or 1000 L. In other embodiments,devices for processing samples accept samples of masses of at leastabout 1 nanogram (ng), or 10 ng, or 100 ng, or 1 microgram (mg), or 10μg, or 100 μg, or 1 milligram (mg), or 10 mg, or 100 mg, or 1 gram (g),or 2 g, or 3 g, or 4 g, or 5 g, or 6 g, or 7 g, or 8 g, or 9 g, or 10 g,or 100 g, or 1000 g.

A device may perform sample preparation, processing and/or detectionwith the aid of one module or a plurality of modules. For example, adevice may prepare a sample in a first module (e.g., the first module701 of FIG. 7) and run (or perform) an assay on the sample in a second(e.g., the second module 702 of FIG. 7) module separate from the firstmodule.

A device may accept one sample or a plurality of samples. In anembodiment, a system accepts a single sample and prepares, processesand/or detects the single sample. In another embodiment, a systemaccepts a plurality of samples and prepares, processes and/or detectsone or more of the plurality of samples at the same time.

In some embodiments, one or more modules of a device are fluidicallyisolated or hydraulically independent from one another. In anembodiment, the plurality of modules 701-706 of the system 700 are influid isolation with respect to one another. In an example fluidisolation is provided by way of seals, such as fluid or pressure seals.In some cases, such seals are hermetic seals. In other embodiments, oneor modules of a system are fluidically coupled to one another.

In some situations, devices having a plurality of modules are configuredto communicate with one another. For example, a first device having aplurality of modules, such as the device 1000, is in communication withanother device, such as a like or similar device having a plurality ofmodules. In such fashion, two or more devices may communicate with oneanother, such as to facilitate resource sharing.

Processing of a biological sample may include pre-processing (e.g.,preparation of a sample for a subsequent treatment or measurement),processing (e.g., alteration of a sample so that it differs from itsoriginal, or previous, state), and post-processing (e.g., fixing asample, or disposing of all or a portion of a sample or associatedreagents following its measurement or use). A biological sample may bedivided into portions, such as aliquots of a blood or urine sample, orsuch as slicing, mincing, or dividing a tissue sample into two or morepieces. Processing of a biological sample, such as blood sample, mayinclude mixing, stirring, sonication, homogenization, or other treatmentof a sample or of a portion of the sample. Processing of a biologicalsample, such as blood sample, may include centrifugation of a sample ora portion thereof. Processing of a biological sample, such as a bloodsample, may include providing time for components of the sample toseparate or settle, and may include filtration (e.g., passing the sampleor a portion thereof through a filter or membrane). Processing of abiological sample, such as a blood sample, may include allowing orcausing a blood sample to coagulate. Processing of a biological sample,such as blood sample, may include concentration of the sample, or of aportion of the sample (e.g., by sedimentation or centrifugation of ablood sample, or of a solution containing a homogenate of tissue from atissue sample, or with electromagnetic other other beads) to provide apellet and a supernatant. Processing of a biological sample, such asblood sample, may include dilution of a portion of the sample. Dilutionmay be of an entire sample, or of a portion of a sample, includingdilution of a pellet or of a supernatant from sample. A biologicalsample may be diluted with water, or with a saline solution, such as abuffered saline solution. A biological sample may be diluted with asolution which may or may not include a fixative (e.g., formaldehyde,paraformaldehyde, or other agent which cross-links proteins). Abiological sample may be diluted with a solution such that an osmoticgradient is produced between the surrounding solution and the interior,or an interior compartment, of such cells, effective that the cellvolume is altered. For example, where the resulting solutionconcentration following dilution is less than the effectiveconcentration of the interior of a cell, or of an interior cellcompartment, the volume of such a cell will increase (i.e., the cellwill swell). A biological sample may be diluted with a solution whichmay or may not include an osmoticant (such as, for example, glucose,sucrose, or other sugar; salts such as sodium, potassium, ammonium, orother salt; or other osmotically active compound or ingredient). Inembodiments, an osmoticant may be effective to maintain the integrity ofcells in the sample, by, for example, stabilizing or reducing possibleosmotic gradients between the surrounding solution and the interior, oran interior compartment, of such cells. In embodiments, an osmoticantmay be effective to provide or to increase osmotic gradients between thesurrounding solution and the interior, or an interior compartment, ofsuch cells, effective that the cells at least partially collapse (wherethe cellular interior or an interior compartment is less concentratedthan the surrounding solution), or effective that the cells swell (wherethe cellular interior or an interior compartment is more concentratedthan the surrounding solution).

A biological sample may be dyed, or markers or reagents may be added tothe sample, or the sample may be otherwise prepared for detection,visualization, or quantification of the sample, a portion of a sample, acomponent part of a sample, or a portion of a cell or structure within asample. For example, a biological sample may be contacted with asolution containing a dye. A dye may stain or otherwise make visible acell, a portion of a cell, a component inside a cell, or a material ormolecule associated with a cell in a sample. A dye may bind to or bealtered by an element, compound, or other component of a sample; forexample a dye may change color, or otherwise alter one of more of itsproperties, including its optical properties, in response to a change ordifferential in the pH of a solution in which it is present; a dye maychange color, or otherwise alter one of more of its properties,including its optical properties, in response to a change ordifferential in the concentration of an element or compound (e.g.,sodium, calcium, CO₂, glucose, or other ion, element, or compound)present in a solution in which the dye is present. For example, abiological sample may be contacted with a solution containing anantibody or an antibody fragment. For example, a biological sample maybe contacted with a solution that includes particles. Particles added toa biological sample may serve as standards (e.g., may serve as sizestandards, where the size or size distribution of the particles isknown, or as concentration standards, where the number, amount, orconcentration of the particles is known), or may serve as markers (e.g.,where the particles bind or adhere to particular cells or types ofcells, to particular cell markers or cellular compartments, or where theparticles bind to all cells in a sample).

In an example, two rack-type devices like the system 700 of FIG. 7 areprovided. The devices are configured to communicate with one another,such as by way of a direct link (e.g., wired network) or wireless link(e.g., Bluetooth, WiFi). While a first of the two rack-type devicesprocesses a portion of a sample (e.g., blood aliquot), a second of thetwo-rack-type devices performs sample detection on another portion ofthe same sample. The first rack-type device then transmits its resultsto the second rack-type device, which uploads the information to aserver in network communication with the second rack-type device but notthe first rack-type device.

Devices and methods provided herein are configured for use with point ofservice systems. In an example, devices are deployable at locations ofhealthcare providers (e.g., drug stores, doctors' offices, clinics,hospitals) for sample preparation, processing and/or detection. In somesituations, devices provided herein are configured for sample collectionand preparation only, and processing (e.g., detection) and/or diagnosisis performed at a remote location certified by a certifying or licensingentity (e.g., government certification).

In some embodiments, a user provides a sample to a system having one ormore modules, such as the system 700 of FIG. 7. The user provides thesample to a sample collection module of the system. In an embodiment,the sample collection module includes one or more of a lancet, needle,microneedle, venous draw, scalpel, cup, swab, wash, bucket, basket, kit,permeable matrix, or any other sample collection mechanism or methoddescribed elsewhere herein. Next, the system directs the sample from thesample collection module to one or more processing modules (e.g.,modules 701-706) for sample preparation, assaying and/or detection. Inan embodiment, the sample is directed from the collection module to theone or more processing modules with the aid of a sample handling system,such as a pipette. Next, the sample is processed in the one or moremodules. In some situations, the sample is assayed in the one or moremodules and subsequently put through one or more detection routines.

In some embodiments, following processing in the one or more modules,the system communicates the results to a user or a system (e.g., server)in communication with the system. Other systems or users may then accessthe results to aid in treating or diagnosing a subject.

In an embodiment, the system is configured for two-way communicationwith other systems, such as similar or like systems (e.g., a rack, suchas that described in the context of FIG. 7) or other computers systems,including servers.

Devices and methods provided herein, by enabling parallel processing,may advantageously decrease the energy or carbon footprint of point ofservice systems. In some situations, systems, such as the system 700 ofFIG. 7, has a footprint that is at most 10%, or 15%, or 20%, or 25%, or30%, or 35%, or 40%, or 45%, or 50%, or 55%, or 60%, or 65%, or 70%, or75%, or 80%, or 85%, or 90%, or 95%, or 99% that of other point ofservice systems.

In some embodiments, methods are provided for detecting analytes. In anembodiment, a processing routine includes detecting the presence orabsence of an analyte. The processing routine is facilitated with theaid of systems and devices provided herein. In some situations, analytesare associated with biological processes, physiological processes,environmental conditions, sample conditions, disorders, or stages ofdisorders, such as one or more of autoimmune disease, obesity,hypertension, diabetes, neuronal and/or muscular degenerative diseases,cardiac diseases, and endocrine diseases.

In some situations, a device processes one sample at a time. However,systems provided herein are configured for multiplexing sampleprocessing. In an embodiment, a device processes multiple samples at atime, or with overlapping times. In an example, a user provides a sampleto a device having a plurality of modules, such as the system 700 ofFIG. 7. The device then processes the sample with the aid of one or moremodules of the device. In another example, a user provides multiplesamples to a device having a plurality of modules. The device thenprocesses the samples at the same time with the aid of the plurality ofmodules by processing a first sample in a first module while processinga second sample in second module.

The system may process the same type of sample or different types ofsamples. In an embodiment, the system processes one or more portions ofthe same sample at the same time. This may be useful if various assayingand/or detection protocols on the same sample are desired. In anotherembodiment, the system processes different types of samples at the sametime. In an example, the system processes a blood and urine sampleconcurrently in either different modules of the system or a singlemodule having processing stations for processing the blood and urinesamples.

In some embodiments, a method for processing a sample with the aid of apoint of service system, such as the system 700 of FIG. 7, comprisesaccepting testing criteria or parameters and determining a test order orschedule based on the criteria. The testing criteria is accepted from auser, a system in communication with the point of service system, or aserver. The criteria are selectable based on a desired or predeterminedeffect, such as minimizing time, cost, component use, steps, and/orenergy. The point of service system processes the sample per the testorder or schedule. In some situations, a feedback loop (coupled withsensors) enables the point of service system to monitor the progress ofsample processing and maintain or alter the test order or schedule. Inan example, if the system detects that processing is taking longer thanthe predetermined amount of time set forth in the schedule, the systemspeeds up processing or adjusts any parallel processes, such as sampleprocessing in another module of the system. The feedback loop permitsreal-time or pseudo-real time (e.g., cached) monitoring. In somesituations, the feedback loop may provide permit reflex testing, whichmay cause subsequent tests, assays, preparation steps, and/or otherprocesses to be initiated after starting or completing another testand/or assay or sensing one or more parameter. Such subsequent tests,assays, preparation steps, and/or other processes may be initiatedautomatically without any human intervention. Optionally, reflex testingis performed in response to an assay result. Namely by way ofnon-limiting example, if a reflex test is ordered, a cartridge ispre-loaded with reagents for assay A and assay B. Assay A is the primarytest, and assay B is the reflexed test. If the result of assay A ismeets a predefined criteria initiating the reflex test, then assay B isrun with the same sample in the device. The device protocol is plannedto account for the possibility of running the reflex test. Some or allprotocol steps of assay B can be performed before the results for assayA are complete. For example, sample preparation can be completed inadvance on the device. It is possible also to run a reflex test with asecond sample from the patient. In some embodiments, devices and systemsprovided herein may contain components such that multiple differentassays and assay types may be reflex tested with the same device. Insome embodiments, multiple tests of clinical significance may beperformed in a single device provided herein as part of a reflex testingprotocol, where the performance of the same tests with known systems andmethods requires two or more separate devices. Accordingly, systems anddevices provided herein may permit, for example, reflex testing which isfaster and requires less sample than known systems and methods. Inaddition, in some embodiments, for reflex testing with a device providedherein, it is not necessary to know in advance which reflexed testedwill be performed.

In some embodiments, the point of service system may stick to apre-determined test order or schedule based on initial parameters and/ordesired effects. In other embodiments, the schedule and/or test ordermay be modified on the fly. The schedule and/or test order may bemodified based on one or more detected conditions, one or moreadditional processes to run, one or more processes to no longer run, oneor more processes to modify, one or more resource/component utilizationmodifications, one or more detected error or alert condition, one ormore unavailability of a resource and/or component, one or moresubsequent input or sample provided by a user, external data, or anyother reason.

In some examples, one or more additional samples may be provided to adevice after one or more initial samples are provided to the device. Theadditional samples may be from the same subject or different subjects.The additional samples may be the same type of sample as the initialsample or different types of samples (e.g., blood, tissue). Theadditional samples may be provided prior to, concurrently with, and/orsubsequent to processing the one or more initial samples on the device.The same and/or different tests or desired criteria may be provided forthe additional samples, as opposed to one another and/or the initialsamples. The additional samples may be processed in sequence and/or inparallel with the initial samples. The additional samples may use one ormore of the same components as the initial samples, or may use differentcomponents. The additional samples may or may not be requested in viewof one or more detected condition of the initial samples.

In some embodiments, the system accepts a sample with the aid of asample collection module, such as a lancet, scalpel, or fluid collectionvessel. The system then loads or accesses a protocol for performing oneor more processing routines from a plurality of potential processingroutines. In an example, the system loads a centrifugation protocol andcytometry protocol. In some embodiments, the protocol may be loaded froman external device to a sample processing device. Alternatively, theprotocol may already be on the sample processing device. The protocolmay be generated based on one or more desired criteria and/or processingroutines. In one example, generating a protocol may include generating alist of one or more subtasks for each of the input processes. In someembodiments, each subtask is to be performed by a single component ofthe one or more devices. Generating a protocol may also includegenerating the order of the list, the timing and/or allocating one ormore resources.

In an embodiment, a protocol provides processing details orspecifications that are specific to a sample or a component in thesample. For instance, a centrifugation protocol may include rotationalvelocity and processing time that is suited to a predetermined sampledensity, which enables density-dependent separation of a sample fromother material that may be present with a desirable component of thesample.

A protocol is included in the system, such as in a protocol repositoryof the system, or retrieved from another system, such as a database, incommunication with the system. In an embodiment, the system is inone-way communication with a database server that provides protocols tothe system upon request from the system for one or more processingprotocols. In another embodiment, the system is in two-way communicationwith a database server, which enables the system to upload user-specificprocessing routines to the database server for future use by the user orother users that may have use for the user-specific processing routines.

In some cases, a processing protocol is adjustable by a user. In anembodiment, a user may generate a processing protocol with the aid of aprotocol engine that provides the user one or more options geared towardtailoring the protocol for a particular use. The tailoring may occurprior to use of the protocol. In some embodiments, the protocol may bemodified or updated while the protocol is in use.

With the aid of a protocol, a system processes a sample, which mayinclude preparing the sample, assaying the sample and detecting one ormore components of interest in the sample. In some cases, the systemperforms data analysis with respect to the sample or a plurality ofsample after processing. In other cases, the system performs dataanalysis during processing. In some embodiments, data analysis isperformed on-board—that is, on the system. In other embodiments, dataanalysis is performed using a data analysis system that is external tothe system. In such a case, data is directed to the analysis systemwhile the sample is being processed or following processing.

In some embodiments, a single sample from a subject provided to a deviceor component thereof may be used for two or more assays. The assays maybe any assays described elsewhere herein. In some embodiments, a sampleprovided to a device may be whole blood. The whole blood may contain ananticoagulant (e.g. EDTA, Coumadins, heparin, or others). Within thedevice, whole blood may be subjected to a procedure to separate bloodcells from plasma (e.g. by centrifugation or filtration). In analternative, a sample containing separated blood cells and plasma may beintroduced into a device (e.g. if a whole blood sample is separated intoplasma and blood cells before insertion of the sample into the device).Whole blood may be used for one or more assays; in such circumstance,the whole blood may be processed (e.g. diluted) prior to the assays. Asample containing plasma and cell-containing portions may be furtherprocessed to prepare one or both of the portions for assays. Forexample, the plasma may be removed from the cells into a new vessel anddiluted with one or more different diluents to generate one or moredifferent sample dilution levels. The plasma samples (diluted ornon-diluted) may be used for one or more different assays, including,for example immunoassays, general chemistry assays, and nucleic acidassays. In some examples, a plasma sample from an original whole bloodsample may be used for at 1, 2, 3, 4, 5, or more immunoassays, 1, 2, 3,4, 5, or more general chemistry assays, and 1, 2, 3, 4, 5, or morenucleic acid assays. In some examples, the plasma samples may be usedfor two or more different assays that result in two or more differentoptical properties that may be measured (for example, an assay mayresult in a change in the color of the assay, a change in the absorbanceof the assay, a change in the turbidity of the assay, a change in thefluorescence of the assay, or a change in luminescence in the assay,etc.). In addition, the cells isolated from the same whole blood sampledescribed above may also be used for one or more assays. For example,the cells may be measured by cytometry. Cytometry assays may include anydescriptions of cytometry provided elsewhere herein, including cellimaging by microscopy and flow cytometry. In some embodiments, cellswhich are centrifuged or otherwise processed in a sub-optimalanticoagulant or other buffer, reagent, or sample condition may still beused for cytometry. In such circumstances, it may be advantageous toseparate the cells rapidly from the sub-optimal conditions (e.g. bycentrifugation or filtration) to minimize the time the cells are exposedto the sub-optimal conditions. In some embodiments, cells are furtherprocessed to separate the cells into different cell fractions or celltypes—e.g. to separate red blood cells from white blood cells. Inaddition, cells may be measured by other types of assays, such asgeneral chemistry assays (e.g. to perform hemagglutination assays forred blood cell typing).

In some embodiments, methods are provided for performing with a devicedescribed herein two or more assays with a single sample from a subject,including one or more of: 1) if the sample is whole blood, separatingthe whole blood into plasma and cell portions, and optionally, retainingsome blood as whole blood; 2) dividing an original sample of whole bloodinto 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 50, 60, 70,80, 100, 200, 300, 400, 500, or more fluidically isolated aliquots; 3)dividing an original sample of plasma into 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 15, 20, 25, 30, 35, 40, 50, 60, 70, 80, 100, 200, 300, 400, 500, ormore fluidically isolated aliquots; 4) diluting an original sample ofplasma into one or more plasma samples having different dilution levels;5) with plasma samples, performing at least one, two, or three assays ofeach of one, two, or three types of assays, the assay types selectedfrom immunoassays, general chemistry assays, and nucleic acid assays; 6)with plasma sample assays, measuring assay results at least one, two, orthree different detection units, such as, photodiodes PMTs,electrodiodes, spectrophotometers, imaging devices, cameras, CCDsensors, and nucleic acid assay station containing a light source and anoptical sensor; 7) separating blood cells into white blood cell or redblood cell containing portions; 8) with cell-containing samples,performing at least one, two, or three assays of each of one, two,three, or four types of assays, the assay types selected fromimmunoassays, general chemistry assays, nucleic acid assays, andcytometry assays; 9) with cytometry assays, obtaining a digital image ofone or more cells; 10) with cytometry assays, obtaining a cell count;11) with cytometry assays, performing flow cytometry and obtainingscatter plots; 12) heating a sample; and 13) processing a sample withany reagent or chemical disclosed elsewhere herein.

Accuracy, Sensitivity, Precision and Coefficient of Variation

Accuracy is the degree of veracity. Precision is the degree ofreproducibility. Accuracy is a measure of a closeness of a measurementto a predetermined target measurement, result, or reference (e.g.,reference value). Precision is the closeness of a multiple measurementsto one another. In some cases, precision is quantified using a meandegree of reproducibility. Accuracy may be quantified using a deviationor spread in relation to a predetermined value.

In some embodiments, the system has a sensitivity that is the sameirrespective of the type of sample being processed. In some instances,the system may be capable of detecting analytes or signals within therange of about one molecule (e.g., nucleic acid molecule), 5 molecules,10 molecules, or within about 1 pg/mL, 5 pg/mL, 10 pg/mL, 50 pg/mL, 100pg/mL, 500 pg/mL, 1 ng/mL, 5 ng/mL, 10 ng/mL, 50 ng/mL, 100 ng/mL, 150ng/mL, 200 ng/mL, 300 ng/mL, 500 ng/mL, 750 ng/mL, 1 μg/mL, 5 μg/mL, 10μg/mL, 50 μg/mL, 100 μg/mL, 150 μg/mL, 200 μg/mL, 300 μg/mL, 500 μg/mL,750 μg/mL, 1 mg/mL, 1.5 mg/mL, 2 mg/mL, 3 mg/mL, 4 mg/mL, 5 mg/mL, 7mg/mL, 10 mg/mL, 20 mg/mL, or 50 mg/mL. In some embodiments, a system,including one or more modules of the system, has a sensitivity that issample-specific. That is, the sensitivity for detection of the system isdependent on one or more parameters that are specific to the sample,such as the type of sample.

In some embodiments, the system has an accuracy that is the sameirrespective of at least one sample parameter that is specific to asample, such as the type of sample. In an embodiment, the system has anaccuracy of at least about 20%, or 25%, or 30%, or 35%, or 40%, or 45%,55%, or 60%, or 65%, or 70%, or 75%, or 80%, or 85%, or 90%, or 91%, or92%, or 93%, or 94%, or 95%, or 96%, or 97%, or 98%, or 99%, or 99.9%,or 99.99%, or 99.999%. The modules, and/or components may have anyaccuracy, including those described elsewhere herein. In someembodiments, a system, including one or more modules of the system, hasan accuracy that is sample-specific. That is, the accuracy of the systemis dependent on at least one sample parameter that is specific to thesample, such as the type of sample. In such a case, the system may beable to provide more accurate results for one type of sample thananother type of sample.

In some embodiments, the system has a precision that is the sameirrespective of at least one parameter that is specific to a sample,such as the type of sample. In other embodiments, the system has aprecision that is sample-specific. In such a case, the system processesone type of sample at a higher precision than another type of sample.

A coefficient of variation is the ratio between the standard deviationand an absolute value of the mean. In an embodiment, the system has acoefficient of variation (CV) (also “relative standard deviation”herein) less than or equal to about 20%, 15%, 12%, 10%, 9%, 8%, 7%, 6%,5%, 4%, 3%, 2%, 1%, 0.5%, 0.3%, or 0.1%. In another embodiment, a modulein the system has a coefficient of variation less than or equal to about20%, 15%, 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.3%, or0.1%. In another embodiment, a processing routine has a coefficient ofvariation less than or equal to about 20%, 15%, 12%, 10%, 9%, 8%, 7%,6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.3%, or 0.1%.

Systems provided herein have coefficients of variation that are suitedfor longitudinal trend analysis, such as research study that involvesrepeated observations of the same variables over a predetermined periodof time. In an example, results from a sample processed with a firstsystem having a CV less than about 15% and a second system having a CVless than about 15% may be correlated to assess trends in health ortreatment of a subject.

Systems provided herein have dynamic ranges suited to processing sampleshaving concentrations ranging over 100 orders of magnitude or more, 50orders of magnitude or more, 30 orders of magnitude or more, 10 ordersof magnitude or more, 7 orders of magnitude or more, 5 orders ofmagnitude or more, 4 orders of magnitude or more, 3 orders of magnitudeor more, 2 orders of magnitude or more, or one order of magnitude ormore. In an example, a system processes the same sample twice, firstwith a sample volume of about 0.1 mL and second with a sample volume ofabout 10 mL. The results of both cases fall within the accuracy,precision and coefficient of variation described above. In addition,systems provided herein are configured to detect signals within a range(“dynamic range”) of over 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20, or more orders of magnitude. In some cases, thedynamic range is enabled by dilution. In an embodiment, dynamic feedbackis used to determine the level of sample dilution.

Sample Processing Rates

In an embodiment, a point of service system or one or more moduleswithin the system is configured to centrifuge a sample in a time periodof at most about 4 hours, or 3 hours, or 2 hours, or 1 hour, or 45minutes, or 30 minutes, or 15 minutes, or 10 minutes, or 9 minutes, or 8minutes, or 7 minutes, or 6 minutes, or 5 minutes, or 4 minutes, or 3minutes, or 2 minutes, or 1 minute, or 45 seconds, or 30 seconds, or 20seconds, or 10 seconds, or 5 seconds, or 3 seconds, or 1 second, or 0.5second, or 0.1 second. In another embodiment, a point of service systemor one or more modules within the system is configured to perform acytometry assay on a sample in a time period of at most about 4 hours,or 3 hours, or 2 hours, or 1 hour, or 45 minutes, or 30 minutes, or 15minutes, or 10 minutes, or 9 minutes, or 8 minutes, or 7 minutes, or 6minutes, or 5 minutes, or 4 minutes, or 3 minutes, or 2 minutes, or 1minute, or 45 seconds, or 30 seconds, or 20 seconds, or 10 seconds, or 5seconds, or 3 seconds, or 1 second, or 0.5 second, or 0.1 second. Inanother embodiment, a point of service system or one or more moduleswithin the system is configured to perform an immunoassay on a sample ina time period of at most about 4 hours, or 3 hours, or 2 hours, or 1hour, or 45 minutes, or 30 minutes, or 15 minutes, or 10 minutes, or 9minutes, or 8 minutes, or 7 minutes, or 6 minutes, or 5 minutes, or 4minutes, or 3 minutes, or 2 minutes, or 1 minute, or 45 seconds, or 30seconds, or 20 seconds, or 10 seconds, or 5 seconds, or 3 seconds, or 1second, or 0.5 second, or 0.1 second. In another embodiment, a point ofservice system or one or more modules within the system is configured toperform a nucleic acid assay on a sample in a time period of at mostabout 4 hours, or 3 hours, or 2 hours, or 1 hour, or 45 minutes, or 30minutes, or 15 minutes, or 10 minutes, or 9 minutes, or 8 minutes, or 7minutes, or 6 minutes, or 5 minutes, or 4 minutes, or 3 minutes, or 2minutes, or 1 minute, or 45 seconds, or 30 seconds, or 20 seconds, or 10seconds, or 5 seconds, or 3 seconds, or 1 second, or 0.5 second, or 0.1second. In another embodiment, a point of service system or one or moremodules within the system is configured to perform a receptor-basedassay on a sample in a time period of at most about 4 hours, or 3 hours,or 2 hours, or 1 hour, or 45 minutes, or 30 minutes, or 15 minutes, or10 minutes, or 9 minutes, or 8 minutes, or 7 minutes, or 6 minutes, or 5minutes, or 4 minutes, or 3 minutes, or 2 minutes, or 1 minute, or 45seconds, or 30 seconds, or 20 seconds, or 10 seconds, or 5 seconds, or 3seconds, or 1 second, or 0.5 second, or 0.1 second. In anotherembodiment, a point of service system or one or more modules within thesystem is configured to perform a colorimetric assay on a sample in atime period of at most about 4 hours, or 3 hours, or 2 hours, or 1 hour,or 45 minutes, or 30 minutes, or 15 minutes, or 10 minutes, or 9minutes, or 8 minutes, or 7 minutes, or 6 minutes, or 5 minutes, or 4minutes, or 3 minutes, or 2 minutes, or 1 minute, or 45 seconds, or 30seconds, or 20 seconds, or 10 seconds, or 5 seconds, or 3 seconds, or 1second, or 0.5 second, or 0.1 second. In another embodiment, a point ofservice system or one or more modules within the system is configured toperform an enzymatic assay on a sample in a time period of at most about4 hours, or 3 hours, or 2 hours, or 1 hour, or 45 minutes, or 30minutes, or 15 minutes, or 10 minutes, or 9 minutes, or 8 minutes, or 7minutes, or 6 minutes, or 5 minutes, or 4 minutes, or 3 minutes, or 2minutes, or 1 minute, or 45 seconds, or 30 seconds, or 20 seconds, or 10seconds, or 5 seconds, or 3 seconds, or 1 second, or 0.5 second, or 0.1second. In another embodiment, a point of service system or one or moremodules within the system is configured to perform a mass spectrometry(or mass spectroscopy) assay on a sample in a time period of at mostabout 4 hours, or 3 hours, or 2 hours, or 1 hour, or 45 minutes, or 30minutes, or 15 minutes, or 10 minutes, or 9 minutes, or 8 minutes, or 7minutes, or 6 minutes, or 5 minutes, or 4 minutes, or 3 minutes, or 2minutes, or 1 minute, or 45 seconds, or 30 seconds, or 20 seconds, or 10seconds, or 5 seconds, or 3 seconds, or 1 second, or 0.5 second, or 0.1second. In another embodiment, a point of service system or one or moremodules within the system is configured to perform an infraredspectroscopy assay on a sample in a time period of at most about 4hours, or 3 hours, or 2 hours, or 1 hour, or 45 minutes, or 30 minutes,or 15 minutes, or 10 minutes, or 9 minutes, or 8 minutes, or 7 minutes,or 6 minutes, or 5 minutes, or 4 minutes, or 3 minutes, or 2 minutes, or1 minute, or 45 seconds, or 30 seconds, or 20 seconds, or 10 seconds, or5 seconds, or 3 seconds, or 1 second, or 0.5 second, or 0.1 second. Inanother embodiment, a point of service system or one or more moduleswithin the system is configured to perform an x-ray photoelectronspectroscopy assay on a sample in a time period of at most about 4hours, or 3 hours, or 2 hours, or 1 hour, or 45 minutes, or 30 minutes,or 15 minutes, or 10 minutes, or 9 minutes, or 8 minutes, or 7 minutes,or 6 minutes, or 5 minutes, or 4 minutes, or 3 minutes, or 2 minutes, or1 minute, or 45 seconds, or 30 seconds, or 20 seconds, or 10 seconds, or5 seconds, or 3 seconds, or 1 second, or 0.5 second, or 0.1 second. Inanother embodiment, a point of service system or one or more moduleswithin the system is configured to perform an electrophoresis assay on asample in a time period of at most about 4 hours, or 3 hours, or 2hours, or 1 hour, or 45 minutes, or 30 minutes, or 15 minutes, or 10minutes, or 9 minutes, or 8 minutes, or 7 minutes, or 6 minutes, or 5minutes, or 4 minutes, or 3 minutes, or 2 minutes, or 1 minute, or 45seconds, or 30 seconds, or 20 seconds, or 10 seconds, or 5 seconds, or 3seconds, or 1 second, or 0.5 second, or 0.1 second. In anotherembodiment, a point of service system or one or more modules within thesystem is configured to perform a nucleic acid sequencing (e.g.,single-molecule sequencing) assay on a sample in a time period of atmost about 4 hours, or 3 hours, or 2 hours, or 1 hour, or 45 minutes, or30 minutes, or 15 minutes, or 10 minutes, or 9 minutes, or 8 minutes, or7 minutes, or 6 minutes, or 5 minutes, or 4 minutes, or 3 minutes, or 2minutes, or 1 minute, or 45 seconds, or 30 seconds, or 20 seconds, or 10seconds, or 5 seconds, or 3 seconds, or 1 second, or 0.5 second, or 0.1second. In another embodiment, a point of service system or one or moremodules within the system is configured to perform an agglutinationassay on a sample in a time period of at most about 4 hours, or 3 hours,or 2 hours, or 1 hour, or 45 minutes, or 30 minutes, or 15 minutes, or10 minutes, or 9 minutes, or 8 minutes, or 7 minutes, or 6 minutes, or 5minutes, or 4 minutes, or 3 minutes, or 2 minutes, or 1 minute, or 45seconds, or 30 seconds, or 20 seconds, or 10 seconds, or 5 seconds, or 3seconds, or 1 second, or 0.5 second, or 0.1 second. In anotherembodiment, a point of service system or one or more modules within thesystem is configured to perform a chromatography assay on a sample in atime period of at most about 4 hours, or 3 hours, or 2 hours, or 1 hour,or 45 minutes, or 30 minutes, or 15 minutes, or 10 minutes, or 9minutes, or 8 minutes, or 7 minutes, or 6 minutes, or 5 minutes, or 4minutes, or 3 minutes, or 2 minutes, or 1 minute, or 45 seconds, or 30seconds, or 20 seconds, or 10 seconds, or 5 seconds, or 3 seconds, or 1second, or 0.5 second, or 0.1 second. In another embodiment, a point ofservice system or one or more modules within the system is configured toperform a coagulation assay on a sample in a time period of at mostabout 4 hours, or 3 hours, or 2 hours, or 1 hour, or 45 minutes, or 30minutes, or 15 minutes, or 10 minutes, or 9 minutes, or 8 minutes, or 7minutes, or 6 minutes, or 5 minutes, or 4 minutes, or 3 minutes, or 2minutes, or 1 minute, or 45 seconds, or 30 seconds, or 20 seconds, or 10seconds, or 5 seconds, or 3 seconds, or 1 second, or 0.5 second, or 0.1second. In another embodiment, a point of service system or one or moremodules within the system is configured to perform electrochemicalmeasurements on a sample in a time period of at most about 4 hours, or 3hours, or 2 hours, or 1 hour, or 45 minutes, or 30 minutes, or 15minutes, or 10 minutes, or 9 minutes, or 8 minutes, or 7 minutes, or 6minutes, or 5 minutes, or 4 minutes, or 3 minutes, or 2 minutes, or 1minute, or 45 seconds, or 30 seconds, or 20 seconds, or 10 seconds, or 5seconds, or 3 seconds, or 1 second, or 0.5 second, or 0.1 second. Inanother embodiment, a point of service system or one or more moduleswithin the system is configured to perform a histology assay on a samplein a time period of at most about 4 hours, or 3 hours, or 2 hours, or 1hour, or 45 minutes, or 30 minutes, or 15 minutes, or 10 minutes, or 9minutes, or 8 minutes, or 7 minutes, or 6 minutes, or 5 minutes, or 4minutes, or 3 minutes, or 2 minutes, or 1 minute, or 45 seconds, or 30seconds, or 20 seconds, or 10 seconds, or 5 seconds, or 3 seconds, or 1second, or 0.5 second, or 0.1 second. In another embodiment, a point ofservice system or one or more modules within the system is configured toperform a live cell analysis (assay) on a sample in a time period of atmost about 4 hours, or 3 hours, or 2 hours, or 1 hour, or 45 minutes, or30 minutes, or 15 minutes, or 10 minutes, or 9 minutes, or 8 minutes, or7 minutes, or 6 minutes, or 5 minutes, or 4 minutes, or 3 minutes, or 2minutes, or 1 minute, or 45 seconds, or 30 seconds, or 20 seconds, or 10seconds, or 5 seconds, or 3 seconds, or 1 second, or 0.5 second, or 0.1second.

In an embodiment, a processing system, such as a point of servicesystem, is configured to perform any one assay selected from the groupconsisting of immunoassay, nucleic acid assay, receptor-based assay,cytometric assay, colorimetric assay, enzymatic assay, electrophoreticassay, electrochemical assay, spectroscopic assay, chromatographicassay, microscopic assay, topographic assay, calorimetric assay,turbidimetric assay, agglutination assay, radioisotope assay,viscometric assay, coagulation assay, clotting time assay, proteinsynthesis assay, histological assay, culture assay, osmolarity assay,and/or other types of assays or combinations thereof in a time period ofat most about 2 hours, or 1 hour, or 30 minutes, or 10 minutes, or 5minutes, or 1 minute, or 30 seconds. In another embodiment, a processingsystem, such as a point of service system, is configured to perform anytwo assays selected from the group consisting of immunoassay, nucleicacid assay, receptor-based assay, cytometric assay, colorimetric assay,enzymatic assay, electrophoretic assay, electrochemical assay,spectroscopic assay, chromatographic assay, microscopic assay,topographic assay, calorimetric assay, turbidimetric assay,agglutination assay, radioisotope assay, viscometric assay, coagulationassay, clotting time assay, protein synthesis assay, histological assay,culture assay, osmolarity assay, and/or other types of assays orcombinations thereof in a time period of at most about 2 hours, or 1hour, or 30 minutes, or 10 minutes, or 5 minutes, or 1 minute. Inanother embodiment, a processing system, such as a point of servicesystem, is configured to perform any three assays selected from thegroup consisting of immunoassay, nucleic acid assay, receptor-basedassay, cytometric assay, colorimetric assay, enzymatic assay,electrophoretic assay, electrochemical assay, spectroscopic assay,chromatographic assay, microscopic assay, topographic assay,calorimetric assay, turbidimetric assay, agglutination assay,radioisotope assay, viscometric assay, coagulation assay, clotting timeassay, protein synthesis assay, histological assay, culture assay,osmolarity assay, and/or other types of assays or combinations thereofin a time period of at most about 3 hours, or 2 hours, or 1 hour, or 30minutes, or 10 minutes, or 5 minutes, or 1 minute.

In an embodiment, a point of service system, such as the system 700 ofFIG. 7, is configured to process at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or10 samples in a time period of at most about 5 hours, or 4 hours, or 3hours, or 2 hours, or 1 hour, or 30 minutes, or 10 minutes, or 5minutes, or 1 minute, or 30 seconds. In another embodiment, a pluralityof point of service systems working in parallel are configured toprocess at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 samples in a timeperiod of at most about 5 hours, or 4 hours, or 3 hours, or 2 hours, or1 hour, or 30 minutes, or 10 minutes, or 5 minutes, or 1 minute, or 30seconds.

In an embodiment, a processing system, such as a point of servicesystem, is configured to collect a sample and processes the sample in atime period of at most about 5 hours, or 4 hours, or 3 hours, or 2hours, or 1 hour, or 30 minutes, or 10 minutes, or 5 minutes, or 1minute, or 30 seconds. In another embodiment, a processing system, suchas a point of service system, is configured to collect a sample,processes the sample and provide (or transmit) results of the processingin a time period of at most about 5 hours, or 4 hours, or 3 hours, or 2hours, or 1 hour, or 30 minutes, or 10 minutes, or 5 minutes, or 1minute, or 30 seconds.

In an embodiment, a processing system, such as a point of servicesystem, is configured to collect a plurality of samples and processesthe samples in a time period of at most about 5 hours, or 4 hours, or 3hours, or 2 hours, or 1 hour, or 30 minutes, or 10 minutes, or 5minutes, or 1 minute, or 30 seconds. In another embodiment, a processingsystem, such as a point of service system, is configured to collect aplurality of samples, processes the samples and provide (or transmit)results of the processing in a time period of at most about 5 hours, or4 hours, or 3 hours, or 2 hours, or 1 hour, or 30 minutes, or 10minutes, or 5 minutes, or 1 minute, or 30 seconds.

In an embodiment, a processing system, such as a point of servicesystem, is configured to collect a sample and assay the sample in a timeperiod of at most about 5 hours, or 4 hours, or 3 hours, or 2 hours, or1 hour, or 30 minutes, or 10 minutes, or 5 minutes, or 1 minute, or 30seconds. In another embodiment, a processing system, such as a point ofservice system, is configured to collect a sample, assay the sample andprovide (or transmit) results of the assaying in a time period of atmost about 5 hours, or 4 hours, or 3 hours, or 2 hours, or 1 hour, or 30minutes, or 10 minutes, or 5 minutes, or 1 minute, or 30 seconds.

In an embodiment, a processing system, such as a point of servicesystem, is configured to collect a sample, prepare the sample and assaythe sample in a time period of at most about 5 hours, or 4 hours, or 3hours, or 2 hours, or 1 hour, or 30 minutes, or 10 minutes, or 5minutes, or 1 minute, or 30 seconds. In another embodiment, a processingsystem, such as a point of service system, is configured to collect asample, prepare the sample, assay the sample and provide (or transmit)results of the assaying in a time period of at most about 5 hours, or 4hours, or 3 hours, or 2 hours, or 1 hour, or 30 minutes, or 10 minutes,or 5 minutes, or 1 minute, or 30 seconds.

In an embodiment, a processing system, such as a point of servicesystem, is configured to collect a sample and perform multiple assays onthe sample in a time period of at most about 5 hours, or 4 hours, or 3hours, or 2 hours, or 1 hour, or 30 minutes, or 10 minutes, or 5minutes, or 1 minute, or 30 seconds. In another embodiment, a processingsystem, such as a point of service system, is configured to collect asample, perform multiple assays on the sample and provide (or transmit)results of the assaying in a time period of at most about 5 hours, or 4hours, or 3 hours, or 2 hours, or 1 hour, or 30 minutes, or 10 minutes,or 5 minutes, or 1 minute, or 30 seconds.

In an embodiment, a processing system, such as a point of servicesystem, is configured to collect a plurality of samples and performmultiple assays on the samples in a time period of at most about 5hours, or 4 hours, or 3 hours, or 2 hours, or 1 hour, or 30 minutes, or10 minutes, or 5 minutes, or 1 minute, or 30 seconds. In anotherembodiment, a processing system, such as a point of service system, isconfigured to collect a plurality of samples, perform multiple assays onthe samples and provide (or transmit) results of the assaying in a timeperiod of at most about 5 hours, or 4 hours, or 3 hours, or 2 hours, or1 hour, or 30 minutes, or 10 minutes, or 5 minutes, or 1 minute, or 30seconds.

A processing system, such as a point of service system, may beconfigured to collect one or more samples and sequence a geneticsignature from the sample. The entire genome may be sequenced orselected portions of the genome may be sequenced. The processing systemmay be configured to collect and sequence the sample in a time period ofat most about 48 hours, 36 hours, 24 hours, 18 hours, 12 hours, 8 hours,6 hours, 5 hours, or 4 hours, or 3 hours, or 2 hours, or 1 hour, or 30minutes, or 10 minutes, or 5 minutes, or 1 minute, or 30 seconds. Inanother embodiment, a processing system, such as a point of servicesystem, is configured to collect a plurality of samples, performmultiple assays on the samples and provide (or transmit) results of theassaying in a time period of at most about 48 hours, 36 hours, 24 hours,18 hours, 12 hours, 8 hours, 6 hours, 5 hours, or 4 hours, or 3 hours,or 2 hours, or 1 hour, or 30 minutes, or 10 minutes, or 5 minutes, or 1minute, or 30 seconds.

Systems provided herein are configured to store data with the aid ofdata storage modules of the system or external storage systems coupledto the systems. In some situations, data collected and/or generatedduring or after sample processing is compressed and storage in aphysical storage medium, such as a hard disk, memory or cache. In anembodiment, data is compressed with the aid of lossless compression.This may minimize or eliminate any loss of data fidelity.

Processing systems described herein are configured for use as point ofservice systems. In an embodiment, a point of service system is a pointof care system. A point of care system may be used at a point of servicelocation, such as a subject's location (e.g., home or business or sportsevent or security screening or combat location), the location of ahealthcare provider (e.g., doctor), a pharmacy or retailer, a clinic, ahospital, an emergency room, a nursing home, a hospice care location, ora laboratory. A retailer may be a pharmacy (e.g., retail pharmacy,clinical pharmacy, hospital pharmacy), drugstore, chain store,supermarket, or grocer. Examples of retailers may include but are notlimited to Walgreen's, CVS Pharmacy, Duane Reade, Walmart, Target, RiteAid, Kroger, Costco, Kaiser Permanente, or Sears. In some situations, apoint of service system (including but not limited to point of caresystem) is deployed at any location that is designated for use by acertifying or licensing entity (e.g., a government certifying entity).In other situations, a point of service system may be used in orembedded in a transportation vehicle, such as a car, boat, truck, bus,airplane, motorcycle, van, traveling medical vehicle, mobile unit,ambulance, fire engine/truck, critical care vehicle, or other vehicleconfigured to transport a subject from one point to another. A samplecollection site may be at a sample acquisition site and/or healthassessment and/or treatment locations (which may include any of thesample collection sites described elsewhere herein including but notlimited to emergency rooms, doctors' offices, urgent care, tents forscreening (which may be in remote locations), a health care professionalwalking into someone's house to provide home care).

The system (device) or a combination of systems (devices) may belocated/positioned at strategic point of service locations. Locationsmay be selected and optimized based on a variety of objectives, such asbut not limited to disease prevalence, rates of disease development,projected disease rates, estimated risk of outbreaks, populationdemographics, government policies and regulations, customer, physicianand patient preferences, access to other technologies at said locations,safety and risk estimates, safety threats, etc. Devices can be relocatedon a periodic basis to improve overall utility on a frequent basis, suchas daily, weekly, monthly, annually, etc. Systems can be updated toimprove performance and/or add functionality. Systems can be updated ona module by module basis. System updates can occur via hardware and/orvia software. Systems can be updated with minimal downtime via featuresenabling blade and/or module extraction and insertion.

Additionally, a point of service location where a sample may becollected from a subject or provided by a subject may be a locationremote to an analyzing laboratory. The sample collection site may have aseparate facility from the laboratory. The sample may or may not becollected fresh from the subject at the point of service location.Alternatively, the sample may be collected from the subject elsewhereand brought to the point of service location. In some embodiments, nosample preparation step is provided on the sample before being providedto the device. For example, no slide needs to be prepped before a sampleis provided to the device. Alternatively, one or more sample preparationstep may be performed on the sample before being provided to the device.

A sample collection site at a point of service location may be a bloodcollection center, or any other bodily fluid collection center. Thesample collection site may be a biological sample collection center. Insome embodiments, a sample collection site may be a retailer. Otherexamples of sample collection sites may include hospitals, clinics,health care professionals' offices, schools, day-care centers, healthcenters, assisted living residences, government offices, travelingmedical care units, or the home. For example, a sample collection sitemay be a subject's home. A sample collection site may be any locationwhere a sample from the subject is received by the device. A collectionsite may be a moving location, such as on or with a patient or in amobile unit or vehicle or with a travelling doctor. Any location may bedesignated as a sample collection site. The designation may be made byany party, including but not limited to the laboratory, entityassociated with the laboratory, governmental agency, or regulatory body.Any description herein relating to sample collection site or point ofservice location may relate to or be applied to retailers, hospitals,clinics, or any other examples provided herein and vice versa.

Point of service systems described in various embodiments, such as apoint of care systems, are configured for with various types of sample,such as, tissue samples (e.g., skin, parts of organs), fluid samples(e.g., breath, blood, urine, saliva, cerebrospinal fluid) and otherbiological samples from a subject (e.g., feces).

Point of service systems described herein are configured to processsamples at a location where the point of service system is accessible bya user. In an example, a point of service system is located at asubject's home and a sample is collected from a subject and processed inthe subject's home. In another example, a point of service system islocated at a drug store and a sample is collected from a subject andprocessed in the drug store. In another example, a point of servicesystem is located at the location of a healthcare provider (e.g.,doctor's office) and a sample is collected from a subject and processedat the location of the healthcare provider. In another example, a pointof service system is located onboard a transportation system (e.g.,vehicle) and a sample is collected from a subject and processed on thetransportation system.

In some embodiments, a sample processing system may be deployed at alocation outside of a central laboratory (e.g. at a school, home, fieldhospital, clinic, business, vehicle, etc.). In some embodiments, asample processing system may be deployed at a location that has aprimary purpose other than laboratory services (e.g. at a school, home,field hospital, clinic, business, vehicle, etc.). In some embodiments,the sample processing system may be deployed at a location that is notdedicated to processing samples received from multiple sampleacquisition locations. In some embodiments, a sample processing systemmay be located less than about 1 kilometer, 500 meters, 400 meters, 300meters, 200 meters, 100 meters, 75 meters, 50 meters, 25 meters, 10meters, 5 meters, 3 meters, 2 meters, or 1 meter from the location atwhich a sample is obtained from a subject. In some embodiments, a sampleprocessing system may be located within the same room, building, orcampus at which a sample is obtained from a subject. In someembodiments, a sample processing system may be on or in a subject. Insome embodiments, a sample may be provided directly from a subject to asample processing system. In some embodiments, a sample may be providedto a sample processing system within 48 hours, 36 hours, 24 hours, 12hours, 8 hours, 6 hours, 4 hours, 3 hours, 2 hours, 1 hour, 45 minutes,30 minutes, 15 minutes, 10 minutes, 5 minutes, 1 minute, or 30 secondsof collection of the sample from a subject.

In some embodiments, a sample processing system may be portable. In someembodiments, a sample processing system may have a total volume of lessthan about 4 m³, 3 m³, 2 m³, 1 m³, 0.5 m³, 0.4 m³, 0.3 m³, 0.2 m³, 0.1m³, 1 cm³, 0.5 cm³, 0.2 cm³, or 0.1 cm³. In some embodiments, a sampleprocessing system may have a mass of than about 1000 kg, 900 kg, 800 kg,700 kg, 600 kg, 500 kg, 400 kg, 300 kg, 200 kg, 100 kg, 75 kg, 50 kg, 25kg, 10 kg, 5 kg, 2 kg, 1 kg, 0.5 kg, 0.1 kg, 25 g, 10 g, 5 g, or 1 g. Insome embodiments, a sample processing system may be configured forambulatory sample processing.

In some embodiments, a system provided herein may be used in space or inzero or low gravity. In such embodiments, the system may be modified toadjust for the reduced gravity, such as by including seals on all liquidvessels and sample handling systems. In addition, steps which rely ongravity (e.g. settling of samples) may instead be achieved by theapplication of a force (e.g. centrifugation). Furthermore, the assaysmay be calibrated to account for changes to the reactions due to lowgravity.

In some embodiments, point of service testing may be performed at thelocation of a sample processing system.

In some embodiments, post-sample processing analysis, includingdiagnosis and/or treatment, is performed by the point of service systemat the location of the point of service system. In other embodiments,post-sample processing analysis is performed remotely from a location inwhich a sample is collected and processed. In an example, post-sampleprocessing analysis is performed at the location of a healthcareprovider. In another example, post-sample processing analysis isperformed at the location of a processing system. In another example,post-sample processing analysis is performed on a server (e.g., on thecloud).

The post-sampling analysis may occur at a laboratory or by an entityaffiliated with a laboratory. A laboratory can be an entity or facilitycapable of performing a clinical test or analyzing collected data. Alaboratory can provide controlled conditions in which scientificresearch, experiments, and measurement can be performed. The laboratorycan be a medical laboratory or clinical laboratory where tests can bedone on clinical specimens, or analysis can occur on data collected fromclinical specimens, in order to get information about the health of apatient as pertaining to the diagnosis, prognosis, treatment, and/orprevention of disease. A clinical specimen may be a sample collectedfrom a subject. Preferably, a clinical specimen may be collected fromthe subject at a sample collection site that is at a separate facilityfrom the laboratory, as described in further detail elsewhere herein.The clinical specimen may be collected from the subject using a device,which is placed at a designated sample collection site or in or on thesubject.

In some embodiments, a laboratory may be a certified laboratory. Thecertified laboratory may be an authorized analytical facility. Anydescription herein of a laboratory may apply to an authorized analyticalfacility and vice versa. In some instances, the laboratory may becertified by a governmental agency. A laboratory may receivecertification or oversight by a regulatory body. In one example, thelaboratory may be certified by an entity, such as Centers for Medicare &Medicaid Services (CMS). For instance, an authorized analytical facilitymay be a Clinical Laboratory Improvement Amendments (CLIA) certifiedlaboratory or its equivalent in any foreign jurisdiction.

In other embodiments, post-processing analysis is performed on thedevice. The same device that receives a sample and/or processes thesample may also perform post-processing analysis. Alternatively thedevice that receives the same and/or processes the sample does notperform post-processing analysis. In some instances, post-processinganalysis may occur both on-board and off-board the device.

In an embodiment, post-processing analysis is performed with the aid ofa post-processing module of the point of service system. In anotherembodiment, post-processing analysis is performed with the aid of apost-processing system that is external to the point of service system.In an example, such post-processing system may be located at ahealthcare provider or other entity that is authorized to performpost-processing analysis.

In some situations, a point of service system is disposed at a locationof a paying party or entity. In an example, a point of service system isdisposed at the location of a healthcare provider that has provided (orwill provide) payment to use the point of service system. In anotherexample, a point of service system is disposed at drugstore that hasprovided (or will provide) payment to use the point of service system.

In an embodiment, post-processing systems enable diagnosis, such asdisease diagnosis. In another embodiment, post-processing systems enabletreatment. In another embodiment, post-processing systems enablediagnosis and treatment. Post-processing systems may be useful fordisease diagnosis, treatment, monitoring, and/or prevention.

In an example, post-processing systems enable drug screening. In such acase, a point of service system collects a sample (e.g., urine sample)from a subject and processes the sample, such as by performingcentrifugation and one or more assays. Next, the point of service systemgenerates data for subsequent post-processing analysis, which includesidentifying (or flagging) whether a predetermined drug has been found inthe sample. The post-processing analysis is done on the system.Alternatively, the post-processing analysis is done at a location remotefrom the location of the point of service system.

In some cases, point of service systems are used in clinical trials,such as for the development of therapeutics. Such clinical trialsinclude one or more procedures conducted to allow safety (or, morespecifically, information about adverse drug reactions and adverseeffects of other treatments) and efficacy data to be collected forhealth interventions (e.g., drugs, diagnostics, devices, therapyprotocols). Point of service systems and information systems providedherein may be used to facilitate enrollment of patients in clinicaltrials, either through testing or through integrated EMR (electronicmedical record) systems or both.

Point of service systems provided herein, in some cases, are configuredfor use in pre-clinical development (or trials). In an example, a pointof service system, such as the system 700 of FIG. 7, is used forprocessing samples and collecting data for feasibility testing,iterative testing and safety, which may be used in pre-clinicaldevelopment. Such trials may include animal testing. Point of servicesystems described herein advantageously enable testing using smallsample volumes at processing rates that enable numerous tests to beperformed with a given sample. Pre-clinical trials with the aid of pointof service systems provided herein enable the assessment of efficacyand/or toxicity of a therapeutic drug or metabolite thereof, or atreatment regimen.

Point of service systems provided herein may optionally be used forbiotoxin testing. The point of service system may process environmentalor product samples, and may detect one or more toxin. Point of servicesystems described herein advantageously enable testing using smallsample volumes at processing rates that enable numerous tests to beperformed with a given sample. Toxin testing with the aid of point ofservice systems provided herein enable the assessment of a threat in theenvironment (e.g., contaminated water, air, soil) or product (e.g., foodand/or beverage products, building materials, and/or any otherproducts).

Point of service systems provided herein, such as the system 700 of FIG.7, enable phylogenetic classification, parental identification, forensicidentification, compliance or non-compliance testing, monitoring adversedrug reactions (ADRs), developing individualized medicine, calibrationof treatment or therapeutic systems and methods, assessing thereliability of treatment or therapeutic systems and methods, and/ortrend analysis (e.g., longitudinal trend analysis). Compliance ornon-compliance testing with the aid of point of service systemsdescribed above may improve patience compliance, which may lowerhealthcare costs associated with complying with a particular treatment.

As part of individualized medicine, a subject uses a point of servicesystem to collect a sample from the subject and process the sample. Inan example, a urine sample is collected from the subject and tested forthe presence of one or more predetermined drugs. In some situations, thecollection of samples, processing of the samples and post-processinganalysis provides subject-specific (or individualized) care. In somecases, following sample collection and processing from a subject, thepoint of service system or post-processing system transmits anotification or alert to the subject or a healthcare provider. In anexample, a point of service system transmits an alert to a subject'sdoctor if the system determines that the concentration of a monitoreddrug (or metabolite of the drug) is above and/or below a predeterminedlimit.

In an embodiment, a point of service system is used to process a sampleand perform post-processing analysis to generate data that is used withother systems. In another embodiment, a point of service system is usedto process a sample and direct post-processing data to another systemfor post-processing analysis with the post-processing data. In such acase, the results of the analysis are configured to be shared with othersystems or individuals, such as if certain access requirements are met.In an example, post-processing data or the results of post-processinganalysis are shared with a payer (e.g., insurance company), healthcareprovider, laboratory, clinic, other point of service device or module,and/or a subject.

Point of service systems may be used to accept, process, and/or analyzea small volume of sample, which may include the volumes describedelsewhere herein. Point of service systems may also be used forproviding rapid results. The point of service systems may be able toprocess and/or analyze a sample within a short amount of time, which mayinclude the lengths of time described elsewhere herein.

Systems provided herein are configured for use as point of servicesystems. Such systems are configured to collect and process one or moresamples at various locations, such as a subject's home or the locationof a healthcare provider. In some embodiments, systems provided herein,such as the system 700 of FIG. 7, have a downtime of at most about 2hours, 1 hour, 30 minutes, 10 minutes, 5 minutes, 1 minute, 30 seconds,1 second, or 0.5 seconds between sample processing routines. In somecases, during the downtime the system resets. In other embodiments,systems provided herein, such as the system 700 of FIG. 7, areconfigured to transmit data to a post-processing system within a timeperiod of at most about 10 minutes, 9 minutes, 8 minutes, 7 minutes, 6minutes, 5 minutes, 4 minutes, 3 minutes, 2 minutes, 1 minute, 30seconds, 10 seconds, 5 seconds, 1 second, 0.5 seconds, 0.1 seconds, or0.01 seconds, or 0.001 seconds after processing. In an example, thesystem 700 collects and processes a first sample and transmits data to apost-processing system. The system 700 is able to accept a second samplefor processing 0.5 seconds after the system 700 transmits data.

In some situations, a system, such as the system 700 of FIG. 7, isconfigured to accept 1, or 2, or 3, or 4, or 5, or 6, or 7, or 8, or 9,or 10 samples per collection routines. In other situations, a system,such as the system 700 of FIG. 7, is configured to accept 1 sample at atime, at a time period of at most about 10 minutes, 9 minutes, 8minutes, 7 minutes, 6 minutes, 5 minutes, 4 minutes, 3 minutes, 2minutes, 1 minute, 30 seconds, 10 seconds, 5 seconds, or 1 secondbetween sample collection points.

In some embodiments, multiple samples may include multiple types ofsamples. In other instances, multiple samples may include the same typeof sample. The multiple samples may be collected from the same subjector from different subjects. The multiple samples may be collected at thesame time or at different points in time. Any combination of these maybe provided for multiple samples.

In some embodiments, point of service systems, such as the system 700 ofFIG. 7, are configured for remote treatment, such as with the aid ofaudio and/or visual media coupled with a communications system, such asa network or telephonic system. In an example, a subject provides asample to a point of service system, which processes the sample togenerate data is processed. Next, the system establishes acommunications link with a remote healthcare provider who reviews thesubject's data and provides a diagnosis. The healthcare provider thenaids the subject in treatment. In an embodiment, the healthcare provideris selected by the subject.

In some embodiments, at least one of the components of the system isconstructed of polymeric materials. Non-limiting examples of polymericmaterials include polystyrene, polycarbonate, polypropylene,polydimethysiloxanes (PDMS), polyurethane, polyvinylchloride (PVC),polysulfone, polymethylmethacrylate (PMMA),acrylonitrile-butadiene-styrene (ABS), and glass.

Systems and subcomponents of the systems may be manufactured by varietyof methods including, without limitation, stamping, injection molding,embossing, casting, blow molding, machining, welding, ultrasonicwelding, and thermal bonding. In an embodiment, a device in manufacturedby injection molding, thermal bonding, and ultrasonic welding. Thesubcomponents of the device may be affixed to each other by thermalbonding, ultrasonic welding, friction fitting (press fitting), adhesivesor, in the case of certain substrates, for example, glass, or semi-rigidand non-rigid polymeric substrates, a natural adhesion between the twocomponents.

Device Use and Identification Methods

The device may be configured to perform only sample processing and datageneration. Alternatively, the device may be configured to performsample processing, data generation as well as subsequent qualitativeand/or quantitative evaluation. In other embodiments, the same devicemay perform sample processing, data generation, and/or qualitativeand/or quantitative evaluation on a case-by-case basis. For example, anycombination of these device functionalities can be applied on a per testbasis, on a per sample basis, on a per patient basis, on a per customerbasis, on a per operator basis, and/or on a per location basis.

Prior to, concurrently with, and/or subsequent to receiving a sample ata device, a subject's identity may be verified. The sample may have beencollected from the subject. A subject's identity may also be verifiedprior to, concurrently with, and/or subsequent to processing a sample ata device. This may include verifying a subject's identity prior to,concurrently with, and/or subsequent to preparing a sample at thedevice, and/or assaying the sample at the device.

In some embodiments, a subject may be associated with a payer. Forexample, a payer, such as a health insurance company, government payer,or any other payer as described herein, may provide coverage for thesubject. A payer may pay some or all of the subject's medical bills. Anydescription herein of the subject's insurance coverage and/or verifyingthe insurance coverage may also apply to any other coverage by anypayer. A subject's insurance coverage may be verified. For example, thesystem may verify that the subject is a member having access toinsurance coverage. The system may also verify that that the subject iseligible for certain tests and/or programs under the insurance. Forexample, certain subjects may be eligible for free diabetes tests orgenetic tests. In some instances, different subjects may be eligible fordifferent tests. Such availability of tests may be customized forindividual subjects or for population groups. Such test eligibility maybe based on a set of rules or guidelines generated for an insurancecompany. Such verification of insurance membership and/or testeligibility may be implemented by a software system.

A subject may arrive at a point of service and may be checked in. Insome embodiments, checking in may include verifying the identity of thesubject. Checking in may also include determining a payer for a subject,such as whether the subject has health insurance coverage. Suchprocedures may be automated at the point of service. The point ofservice may include a physician's office, a retailer site, or any otherpoint of service as described elsewhere herein. In some embodiments, thedevice may be used to check in the subject. Alternatively, an externaldevice which may or may not be in communication with the device may beused to check in the subject. Checking a subject in may permit a systemto access one or more pre-existing records for the subject.

In some embodiments, when a subject arrives at a point of service, theidentification of the subject may be verified. In some embodiments, asample collected from the subject may arrive at a point of service withor without the subject. The identification of the subject may beverified using the device, and/or verified by personnel at the point ofservice. For example, the personnel at the point of service may view thesubject's identification and/or insurance card. The device may or maynot capture an image of the subject and/or collect one or more biometricparameter from the subject. The device may assess one or morecharacteristics associated with the subject including but not limited tosubject's appearance, facial recognition, retinal scan, fingerprintscan, handprint scan, weight, height, circumference, voice, gait,movement, proportions, proteomic data, genetic data, analyte levels,heart rate, blood pressure, electrophysiological readings, and/or bodytemperature, in order to assist with identifying the subject. One ormore of the characteristics of the subject that may be assessed mayinclude one or more physiological parameters of the subject, which mayinclude one or more of the characteristics listed above (e.g., heartrate, blood pressure, electrophysiological readings, body temperature).The device may generate a genetic signature for the subject from asample collected from the subject, and compare the genetic signaturewith a pre-stored genetic signature for the subject. The device may alsogenerate a proteomic signature for the subject from a sample collectedfrom the subject, and compare the proteomic signature with a pre-storedproteomic signature for the subject. In some embodiments, a subject'sidentification may be verified when a genetic signature matches thepre-stored genetic signature. An exact match and/or approximate matchmay be required. A subject's identification may be verified when adifference between the proteomic signature and a pre-stored proteomicsignature falls within an acceptable range. The subject's identificationmay be verified using a combination of a static and dynamic signatureverification from one or more biological sample of the subject. Forexample, a subject's genetic signature may be static while the subject'sproteomic signature may be dynamic. Other examples of dynamic signaturesmay include one or more analyte levels, and/or other physiologicalcharacteristics of the subject.

Identity verification may include comparing one or more static and/ordynamic signature information with previously stored informationrelating to the subject. The previously stored information may beaccessed by the device. The previously stored information may beon-board or external to the device. Identity verification may alsoincorporate general knowledge that need not be subject-specific. Forexample, the verification may flag a possible issue for a dynamicsignature if the subject's height changes drastically when the subjectis a fully grown adult, but may not flag an issue if the subject'sheight changes within an acceptable range when the subject is a growingchild or adolescent. The general knowledge may be on-board or externalto the device. The general knowledge may be stored in one or morememory. In some embodiments, the device and/or an external device may becapable of data mining public information provided across a network.

Verification may occur on-board the device. Alternatively, theidentification of the subject may be collected at the point of serviceand may be further verified at another entity or location. The otherentity or location may verify identity and/or coverage automaticallywithout human intervention, or with human intervention. Verification mayoccur on-board and/or off-board using a software program. In someexamples, a laboratory, health care professional, or payer may verifythe subject identity. The device, laboratory, health care professional,and/or payer may be capable of accessing subject information, such aselectronic health records. Verification may occur rapidly and/or inreal-time. For example, verification may occur within 1 hour or less, 30minutes or less, 20 minutes or less, 15 minutes or less, 10 minutes orless, 5 minutes or less, 3 minutes or less, 1 minute or less, 45 secondsor less, 30 seconds or less, 20 seconds or less, 15 seconds or less, 10seconds or less, 5 seconds or less, 3 seconds or less, 1 second or less,0.5 seconds or less, or 0.1 seconds or less. The verification may beautomated without requiring any human intervention.

The system may verify the identity of the subject for the system'srecords, for insurance coverage, to reduce cost, to save time, toprevent fraud, or any other purpose. The verification may be performedby the device. The verification may be performed by an entity orexternal device in communication with the device. The verification mayoccur at any time. In one example, the subject's identity may beverified prior to preparing the subject's sample for the test. Thesubject's identity may be verified prior to providing a sample to thedevice and/or cartridge. The verification of the subject's identity maybe provided prior to, currently with, or after verifying the subject'sinsurance coverage. The verification of the subject's identity may beprovided prior to, currently with, or after verifying the subject hasreceived a prescription to undergo said qualitative and/or quantitativeevaluation. The verification may take place through communications withthe medical care provider, laboratory, payer, laboratory benefitsmanager, or any other entity. Verification may occur by accessing one ormore data storage units. The data storage units may include anelectronic medical records database and/or a payer database. Anelectronic medical records database may include any information relatingto the subject's health, medical records, history, or treatment.

Verification may occur rapidly and/or in real-time. For example,verification may occur within 10 minutes or less, 5 minutes or less, 3minutes or less, 1 minute or less, 45 seconds or less, 30 seconds orless, 20 seconds or less, 15 seconds or less, 10 seconds or less, 5seconds or less, 3 seconds or less, 1 second or less, 0.5 seconds orless, or 0.1 seconds or less. The verification may be automated withoutrequiring any human intervention.

The verification may include information provided by the subject. Forexample, the verification may include scanning an identification cardand/or insurance card of the subject. The verification may includetaking a picture of the subject and/or the subject's face. For example,the verification may include taking a two-dimensional orthree-dimensional snapshot of the subject. Cameras may be used which mayprovide a two-dimensional digital image of the subject and/or that maybe capable of formulating a three-dimensional or four-dimensional imageof the subject. In some embodiments, a plurality of cameras may be usedsimultaneously. A four-dimensional image of the subject may incorporatechanges over time. The verification may include taking a picture of thesubject's face for identification. The verification may include taking apicture of another portion of the subject's face for identification,including but not limited to the patient's whole body, arm, hand, leg,torso, foot, or any other portion of the body. The verification mayemploy a video camera and/or a microphone that may capture additionalvisual and/or audio information. The verification may include comparingthe subject's movements (e.g., gait), or voice.

The verification of a subject may include entering personal informationrelated to the subject, such as the subject's name, insurance policynumber, answers to key questions, and/or any other information. Theverification may include collecting one or more biometric read-out ofthe subject. For example, the verification may include a fingerprint,handprint, footprint, retinal scan, temperature readout, weight, height,audio information, electrical readouts, or any other information. Thebiometric information may be collected by the device. For example, thedevice may have a touchscreen upon which the subject may put thesubject's palm to be read by the device. The touchscreen may be capableof scanning one or more body part of the subject, and/or receiving atemperature, electrical, and/or pressure readout from the subject.

In some embodiments, the touchscreen may be capable of measuring abody-mass index for the subject. Such a measurement may be based on anelectrical readout from the subject. In one example a method formeasuring the body-fat percentage of a subject may be provided,comprising providing a touchscreen, and placing a first finger on afirst side of the touchscreen and a second finger on a second side ofthe touchscreen. A current may be directed through the body of thesubject, wherein the current is directed through the body of the subjectthrough the first finger and the second finger. The body-fat percentageof the subject may be determined by measuring the resistance between thefirst finger and the second finger with the aid of the current directedthrough the body of the subject. The touchscreen may be a capacitivetouchscreen or resistive touchscreen. In one example, the touchscreenmay be at least a 60-point touchscreen. The first finger may be on afirst hand of the subject and the second finger may be on a second handof the subject. In one non-limiting example, the bioelectrical impedanceanalysis (BIA) method allows one to estimate body fat percentage. Thegeneral principle behind BIA: two or more conductors are attached to aperson's body and a small electric current is sent through the body. Theresistance between the conductors will provide a measure of body fatbetween a pair of electrodes, since the resistance to electricity variesbetween adipose, muscular and skeletal tissue. Calculation of fatpercentage uses the weight, so that must be measured with scales,estimated by the device computer vision system, and/or entered by theuser. Another implementation measures impendence with conductors appliedto both hands and feet for additional accuracy.

Alternatively, the device may receive the biometric information fromother devices. For example, the device may receive the subject's weightfrom a scale that may be separate from the device. The information maybe sent directly from the other devices (e.g., over wired or wirelessconnection) or may be entered manually.

The verification may also include information based on a samplecollected from the subject. For example, the verification may include agenetic signature of the subject. When the sample is provided to thedevice, the device may use at least part of the sample to determine thegenetic signature of the subject. For example, the device may performone or more nucleic acid amplification step and may determine keygenetic markers for the subject. This may form the subject's geneticsignature. The subject's genetic signature may be obtained prior to,concurrently with, or after processing the sample on the device. Thesubject's genetic signature may be stored on one or more data storageunit. For example, the subject's genetic signature may be stored in thesubject's electronic medical records. The subject's collected geneticsignature may be compared with the subject's genetic signature alreadystored in the records, if it exists. Any other unique identifyingcharacteristic of the subject may be used to verify the subject'sidentity.

Methods for the amplification of nucleic acids, including DNA and/orRNA, are known in the art. Amplification methods may involve changes intemperature, such as a heat denaturation step, or may be isothermalprocesses that do not require heat denaturation. The polymerase chainreaction (PCR) uses multiple cycles of denaturation, annealing of primerpairs to opposite strands, and primer extension to exponentiallyincrease copy numbers of the target sequence. Denaturation of annealednucleic acid strands may be achieved by the application of heat,increasing local metal ion concentrations (e.g. U.S. Pat. No.6,277,605), ultrasound radiation (e.g. WO/2000/049176), application ofvoltage (e.g. U.S. Pat. No. 5,527,670, U.S. Pat. No. 6,033,850, U.S.Pat. No. 5,939,291, and U.S. Pat. No. 6,333,157), and application of anelectromagnetic field in combination with primers bound to amagnetically-responsive material (e.g. U.S. Pat. No. 5,545,540), whichare hereby incorporated by reference in their entirety. In a variationcalled RT-PCR, reverse transcriptase (RT) is used to make acomplementary DNA (cDNA) from RNA, and the cDNA is then amplified by PCRto produce multiple copies of DNA (e.g. U.S. Pat. No. 5,322,770 and U.S.Pat. No. 5,310,652, which are hereby incorporated by reference in theirentirety).

One example of an isothermal amplification method is strand displacementamplification, commonly referred to as SDA, which uses cycles ofannealing pairs of primer sequences to opposite strands of a targetsequence, primer extension in the presence of a dNTP to produce a duplexhemiphosphorothioated primer extension product, endonuclease-mediatednicking of a hemimodified restriction endonuclease recognition site, andpolymerase-mediated primer extension from the 3′ end of the nick todisplace an existing strand and produce a strand for the next round ofprimer annealing, nicking and strand displacement, resulting ingeometric amplification of product (e.g. U.S. Pat. No. 5,270,184 andU.S. Pat. No. 5,455,166, which are hereby incorporated by reference intheir entirety). Thermophilic SDA (tSDA) uses thermophilic endonucleasesand polymerases at higher temperatures in essentially the same method(European Pat. No. 0 684 315, which is hereby incorporated by referencein its entirety).

Other amplification methods include rolling circle amplification (RCA)(e.g., Lizardi, “Rolling Circle Replication Reporter Systems,” U.S. Pat.No. 5,854,033); helicase dependent amplification (HDA) (e.g., Kong etal., “Helicase Dependent Amplification Nucleic Acids,” U.S. Pat. Appln.Pub. No. US 2004-0058378 A1); and loop-mediated isothermal amplification(LAMP) (e.g., Notomi et al., “Process for Synthesizing Nucleic Acid,”U.S. Pat. No. 6,410,278), which are hereby incorporated by reference intheir entirety. In some cases, isothermal amplification usestranscription by an RNA polymerase from a promoter sequence, such as maybe incorporated into an oligonucleotide primer. Transcription-basedamplification methods commonly used in the art include nucleic acidsequence based amplification, also referred to as NASBA (e.g. U.S. Pat.No. 5,130,238); methods which rely on the use of an RNA replicase toamplify the probe molecule itself, commonly referred to as Qβ replicase(e.g., Lizardi, P. et al. (1988) BioTechnol. 6, 1197-1202);self-sustained sequence replication (e.g., Guatelli, J. et al. (1990)Proc. Natl. Acad. Sci. USA 87, 1874-1878; Landgren (1993) Trends inGenetics 9, 199-202; and HELEN H. LEE et al., NUCLEIC ACID AMPLIFICATIONTECHNOLOGIES (1997)); and methods for generating additionaltranscription templates (e.g. U.S. Pat. No. 5,480,784 and U.S. Pat. No.5,399,491), which are hereby incorporated by reference in theirentirety. Further methods of isothermal nucleic acid amplificationinclude the use of primers containing non-canonical nucleotides (e.g.uracil or RNA nucleotides) in combination with an enzyme that cleavesnucleic acids at the non-canonical nucleotides (e.g. DNA glycosylase orRNaseH) to expose binding sites for additional primers (e.g. U.S. Pat.No. 6,251,639, U.S. Pat. No. 6,946,251, and U.S. Pat. No. 7,824,890),which are hereby incorporated by reference in their entirety. Isothermalamplification processes can be linear or exponential.

Nucleic acid amplification for subject identification may comprisesequential, parallel, or simultaneous amplification of a plurality ofnucleic acid sequences, such as about or more than about 10, 11, 12, 13,14, 15, 20, 25, 30, 35, 40, 50, 100, or more target sequences. In someembodiments, a subjects entire genome or entire transcriptome isnon-specifically amplified, the products of which are probed for one ormore identifying sequence characteristics. An identifying sequencecharacteristic includes any feature of a nucleic acid sequence that canserve as a basis of differentiation between individuals. In someembodiments, an individual is uniquely identified to a selectedstatistical significance using about or more than about 10, 11, 12, 13,14, 15, 20, 25, 30, 35, 40, 50, 100, or more identifying sequences. Insome embodiments, the statistical significance is about, or smaller thanabout 10⁻², 10⁻³, 10⁻⁴, 10⁻⁵, 10⁻⁶, 10⁻⁷, 10⁻⁸, 10⁻⁹, 10⁻¹⁰, 10⁻¹¹,10⁻¹², 10⁻¹³, 10⁻¹⁴, 10⁻¹⁵ or smaller. Examples of identifying sequencesinclude Restriction Fragment Length Polymorphisms (RFLP; Botstein, etal., Am. J. Hum. Genet. 32: 314-331, 1980; WO 90/13668), SingleNucleotide Polymorphisms (SNPs; Kwok, et al., Genomics 31: 123-126,1996), Randomly Amplified Polymorphic DNA (RAPD; Williams, et al., Nucl.Acids Res. 18: 6531-6535, 1990), Simple Sequence Repeats (SSRs; Zhao &Kochert, Plant Mol. Biol. 21: 607-614, 1993; Zietkiewicz, et al.Genomics 20: 176-183, 1989), Amplified Fragment Length Polymorphisms(AFLP; Vos, et al., Nucl. Acids Res. 21: 4407-4414, 1995), Short TandemRepeats (STRs), Variable Number of Tandem Repeats (VNTR),microsatellites (Tautz, Nucl. Acids. Res. 17: 6463-6471, 1989; Weber andMay, Am. J. Hum. Genet. 44: 388-396, 1989), Inter-RetrotransposonAmplified Polymorphism (IRAP), Long Interspersed Elements (LINE), LongTandem Repeats (LTR), Mobile Elements (ME), RetrotransposonMicrosatellite Amplified Polymorphisms (REMAP), Retrotransposon-BasedInsertion Polymorphisms (RBIP), Short Interspersed Elements (SINE), andSequence Specific Amplified Polymorphism (SSAP). Additional examples ofidentifying sequences are known in the art, for example inUS20030170705, which is incorporated herein by reference. A geneticsignature may consist of multiple identifying sequences of a single type(e.g. SNPs), or may comprise a combination of two or more differenttypes of identifying sequences in any number or combination.

Genetic signatures can be used in any process requiring theidentification of one or more subjects, such as in paternity ormaternity testing, in immigration and inheritance disputes, in breedingtests in animals, in zygosity testing in twins, in tests for inbreedingin humans and animals; in evaluation of transplant suitability such aswith bone marrow transplants; in identification of human and animalremains; in quality control of cultured cells; in forensic testing suchas forensic analysis of semen samples, blood stains, and otherbiological materials; in characterization of the genetic makeup of atumor by testing for loss of heterozygosity; and in determining theallelic frequency of a particular identifying sequence. Samples usefulin the generation of a genetics signature include evidence from a crimescene, blood, blood stains, semen, semen stains, bone, teeth, hair,saliva, urine, feces, fingernails, muscle or other soft tissue,cigarettes, stamps, envelopes, dandruff, fingerprints, items containingany of these, and combinations thereof. In some embodiments, two or moregenetic signatures are generated and compared. In some embodiments, oneor more genetics signatures are compared to one or more known geneticsignatures, such as genetic signatures contained in a database.

The genetic signature may be generated by the device that receives thesample. The genetic signature may be generated by the device thatprepares the sample and/or runs one or more assay. Data collected fromthe device may be sent to an external device that may generate thegenetic signature. The genetic signature may be generated in combinationon the device and an external device.

In some embodiments, a system or device is linked to an automatedLaboratory Automation System (LAS). This Laboratory Automation Systemmay operate on the device or on an external device, including in thecloud. In some embodiments, it may operate in a centralized laboratoryfacility. The Laboratory Automation System may be operably linked to aLaboratory Information System (LIS), which is linked to and, in someembodiments, integrated into an Electronic Medical Records system (EMR).As assay or other data is generated on the device, the data may be fedinto the LAS for analysis and in turn into the LIS and EMR systems. Aself-learning data engine may be integrated into the EMR system suchthat as data is transmitted into the EMR, models running within the EMRcan refit and retune based on real-time data from the field. Such modelsmay power clinical decision support systems which may indexcomprehensive biochemical data generated from a wide range of assaymethodologies running simultaneously on the devices against clinicalrules-based systems, which can be customized in or imported into the EMRsystems databases from various sources, such as hospitals, academiccenters, or laboratories. Algorithms in the self-learning data enginemay power selection of the appropriate clinical decision supportprograms for the end-user. Such selection may happen dynamically basedon the data generated on the device or data prompts entered into thedevice. In some embodiments, certain applications of clinical decisionsupport systems are provided below.

In some embodiments, a system may verify whether the subject hasreceived instruction to undergo a clinical test from a health careprofessional. The system may thus verify whether a subject has receivedan order from a health care professional to undertake a qualitativeand/or quantitative evaluation of a biological sample. For example, thesystem may verify whether the subject has received a prescription fromthe health care professional to take the test. The system may verifywhether the subject has received instructions from the health careprofessional to provide a sample to the device. The system may alsoverify whether the subject was authorized to go to a particular point ofservice to undergo the test. The verification may occur with aid of thedevice. The verification may occur at any time. In one example, thesubject's authorization to take the test may be verified prior topreparing the subject's sample for the test. The subject's authorizationto take the test may be verified prior to providing a sample to thedevice and/or cartridge. The verification of the subject's authorizationmay be provided after verifying the subject's identification. Theverification of the subject's authorization may be provided before orafter verifying the subject has insurance coverage for the clinicaltest. The system may verify whether the subject is covered by healthinsurance for a qualitative and/or quantitative evaluation of a sample,within the verifying step is performed prior to, concurrently with, orafter processing a biological sample with the aid of a device, ortransmitting the data from the device. The verification may take placethrough communications with the medical care provider, laboratory,payer, laboratory benefits manager, or any other entity. Verificationmay occur rapidly and/or in real-time. For example, verification mayoccur within 10 minutes or less, 5 minutes or less, 3 minutes or less, 1minute or less, 45 seconds or less, 30 seconds or less, 20 seconds orless, 15 seconds or less, 10 seconds or less, 5 seconds or less, 3seconds or less, 1 second or less, 0.5 seconds or less, or 0.1 secondsor less. The verification may be automated without requiring any humanintervention.

The system may also verify whether the subject has insurance coverage(and/or coverage by any other payer) for the one or more sampleprocessing steps to occur. The system may verify whether the subject hasinsurance coverage, and also whether the subject has the coverage forthe specific requested tests. The system may verify whether the subjecthas insurance coverage to provide a sample to the device. The system mayalso verify whether the subject has insurance coverage for going to thepoint of service and undergoing one or more test. The verification mayoccur at any time. In one example, the subject's insurance coverage maybe verified prior to preparing the subject's sample for the test. Thesubject's insurance coverage may be verified prior to providing a sampleto the device and/or cartridge. The verification of the subject'sinsurance coverage may be provided after verifying the subject'sidentification. The verification of the subject's insurance coverage maybe provided before or after verifying the subject has received aprescription to take the clinical test. The verification may take placethrough communications with the medical care provider, laboratory,payer, laboratory benefits manager, or any other entity. Theverification may occur with the aid of the device. Verification mayoccur rapidly and/or in real-time. For example, verification may occurwithin 10 minutes or less, 5 minutes or less, 3 minutes or less, 1minute or less, 45 seconds or less, 30 seconds or less, 20 seconds orless, 15 seconds or less, 10 seconds or less, 5 seconds or less, 3seconds or less, 1 second or less, 0.5 seconds or less, or 0.1 secondsor less. The verification may be automated without requiring any humanintervention.

The system may also verify whether the clinical test is appropriate forthe subject. The system may verify whether an order for a qualitativeand/or quantitative evaluation is within a set of policy restrictions.Such policy restrictions may form guidelines. Such policy restrictionsmay be policy restriction of a payer, prescribing physician or otherordering health care professional, laboratory, governmental orregulatory body, or any other entity. Such verification may depend onone or more known characteristic of the subject including but notlimited to gender, age, or past medical history. A clinical decisionsupport system may be provided. The system may be capable of accessingone or more medical records, or information associated with the subject.The system may also be able to access general medical data. The systemmay be able to access records relating to the identity of the subject,insurance coverage of the subject, past and present medical treatmentsof the subject, biological features of the subject, and/or prescriptionsprovided to the subject. The system may be able to access electronichealth records and/or pull up patient records and history. The systemmay also be able to pull up payer records, such as insurance andfinancial information relating to the subject. The verification mayoccur with the aid of the device.

In determining appropriateness of a test, the system may provideadditional front-end decision support. For example, if a physicianordered the same test for the subject the previous week, and it is notthe type of test that needs to be repeated within a week, the system maydetermine that the test is not appropriate. In another example, if thetest somehow conflicts with a previous test or would not be appropriatein view of a treatment the subject is undergoing, the system maydetermine that the test is inappropriate.

In some embodiments, prior to providing a qualitative and/orquantitative evaluation, the system may be capable of accessing one ormore records database and/or payer database. In some instances, thesystem may be capable of determining which records database and/or payerdatabase to access prior to providing said qualitative and/orquantitative evaluation, and/or prior to accessing said databases.Additionally, the system may be capable of accessing general informationthat may or may not be specific to the subject or a peer group of thesubject. The system may be capable of web crawling and/or mining publicinformation, which may include information on a network, such as theInternet. The system may make such determination based on the subject'sidentity, the subject's payer information, information collected aboutthe sample, the proposed qualitative and/or quantitative evaluation,and/or any other information.

In one example, an inappropriate test may be a pregnancy test for a malesubject or a PSA level (prostrate-specific antigen) for a femalesubject. Such tests may fall outside the policy restrictions of a payeror prescribing physician. Such ordering errors may be detectable byreviewing the test ordered and information associated with the subject.Such information associated with the subject may include medical recordsfor the subject or identifying information about the subject. In oneexample, the appropriateness of the test is verified prior to preparingthe subject's sample for the test. The subject's test appropriatenessmay be verified prior to, concurrently with, or subsequent to providinga sample to the device and/or cartridge. The verification of thesubject's test appropriateness may be provided after or prior toverifying the subject's identification and/or insurance coverage. Theverification may take place through communications with the medical careprovider, laboratory, payer, laboratory benefits manager, or any otherentity. A clinical decision support system may operate rapidly and/or inreal-time. For example, verification may occur within 10 minutes orless, 5 minutes or less, 3 minutes or less, 1 minute or less, 45 secondsor less, 30 seconds or less, 20 seconds or less, 15 seconds or less, 10seconds or less, 5 seconds or less, 3 seconds or less, 1 second or less,0.5 seconds or less, or 0.1 seconds or less. The clinical decisionsupport system may be automated without requiring any humanintervention.

In some embodiments, qualified personnel may assist with collecting thesubject's identity and/or providing a sample from the subject to thedevice. The qualified personnel may be an authorized technician that hasbeen trained to use the device. The qualified personnel may be adesignated operator of the device. The qualified personnel may or maynot be a health care professional. In some embodiments, the identity ofthe qualified personnel may be verified. The qualified personnel'sidentity may be verified prior to, currently with, or after receivingthe biological sample, transmitting the data from the deviceelectronically and/or analyzing the transmitted data. The qualifiedpersonnel's identity may be verified prior to, currently with, or afterverifying the identity of the subject. The qualified person's identitymay be verified using one or more of the techniques described elsewhereherein.

The system may be capable of providing one or more laboratory reports.The laboratory reports may be provided to a health care professional. Insome instances, a laboratory report may be provided to a subject. Thelaboratory report may be provided via a user interface on a sampleprocessing device. Alternatively, the laboratory may be provided to oneor more external devices. The laboratory report may include data thatmay be viewed longitudinally. The data may include information collectedover time. Such information over time may include biochemical data,analyte levels, physiological information, lifestyle information,medical care and treatment information, and/or any other informationthat may be collected by a device. One or more graph or chart may showthe change or stability of the information over time. One or moreprojected trend may also be displayed.

In some situations, a laboratory report (or other report of or relatedto the health, condition, or well-being of a subject) is prepared withthe aid of methods (e.g., multivariate methods) provided in U.S. patentapplication Ser. No. 12/412,334 to Michelson et al. (“METHODS ANDSYSTEMS FOR ASSESSING CLINICAL OUTCOMES”), which is entirelyincorporated herein by reference. In an example, a laboratory reportincludes details as to the trajectory, velocity and/or acceleration ofthe progression of a condition (e.g., health or disease condition) of asubject. The trajectory may be indicative of the likelihood ofprogression to the clinical outcome. A laboratory report may be preparedwith the aid of asynchronous data management.

In some embodiments, the longitudinal data may be displayed on thesample processing device. The sample processing device may process asample and transmit data to an external device. Analysis may occurexternal to the device or on-board the device. The result of theanalysis which may include one or more laboratory report, electronicmedical record, laboratory analysis, medical consultation, medicalreference, or any other display, may be displayed on the sampleprocessing device. Any description herein of laboratory report and/orany other item on the aforementioned list may apply to mention of anyother item on the aforementioned list. Alternatively, the laboratoryreport, electronic medical record, or any other display may be displayedon a device external to the sample processing device.

The display of data may include longitudinal data presented over time.Such longitudinal data may account for changes in values, rates ofchanges of values, rates of rates of changes of value, or any furtherrates of change thereof. Such longitudinal data may include predictivedata and/or past estimated data. Such information may include graphicsor charts showing such data over time. Such information include videosthat show change of an image over time. Such data may include evaluativeinformation. Such information may include information relating todiagnosis, prognosis, and/or treatment.

The longitudinal analysis may be possible due to low coefficient ofvariation of the data collected. The longitudinal display and/oranalysis may be based on data having a coefficient of variation havingany of the values described elsewhere herein. In some cases, thelongitudinal analysis may be possible due to high frequency of testing.In some cases, high frequency of testing is enabled by convenient pointof service locations, such as drug stores, doctors' offices, clinics,hospitals, supermarkets, or subjects' homes or offices.

The system may include automated clinical decision support. The clinicaldecision support may include a front-end clinical decision supportsystem and/or a back-end clinical decision support system. In oneexample of a front-end system, when a test is ordered for a subject, theclinical decision support system may indicate whether a test isappropriate/inappropriate for a subject, whether the subject has alreadyundergone the test (e.g., if the test was conducted recently, it mayshow the test results rather than conducing the test), and/or whether asubject is undergoing too many tests. The clinical decision support mayalso recommend additional tests for a subject. In some embodiments, datamay be provided in real-time on a user interface, such as a touchscreen.The displayed data may be customized for an individual viewing the data,or may be customized based on the data. For example, the display andassociated clinical decision support may be customized for a health careprofessional based on biochemical data. A customized health report orthe analysis may display customized recommendations based on bestpractices from relevant clinical decision support systems and providebetter insight into disease onset, progression, and regression, through,e.g., the analysis, longitudinal and other multi-variate (ormultivariate) analyses on the data. The analysis report may includeinformation from the existing EMR system analysis or any results of anytests for a subject described herein, and/or any prognosis or treatmentplans or otherwise health advice tailored for a given subject.

In one example of a back-end system the clinical decision support systemmay refer to one or more guidelines or rules. The guidelines/rules maybe customized per health care professional, per subject, per healthinsurance company or other payer, per hospital, clinic or other medicalentity, or any other group. In some instances, the guidelines/rules maybe customized based on biochemical data. The clinical decision supportsystem may take biochemical data and customize a recommendation for asubject based on lifestyle information, dietary information, or anyother information that may be collected, including those describedelsewhere herein. In some in stances, the back-end clinical decisionsupport may take the data (e.g., including the biochemical data) andcustomize one or more financial transaction. Such financial transactionsmay include reimbursements for an insurance company, and/or health careprofessional, or charging for one or more services.

The clinical decision support may be linked to one or more subject'srecords. The clinical decision support may be linked to the subject'smedical records and/or payer records. The clinical decision support mayintegrate the use of additional general knowledge. The clinical decisionsupport may be updated periodically or continuously to accommodateup-to-date clinical knowledge. The clinical decision supports mayinclude best practices or data associated with diagnosing, treating,monitoring, and/or preventing one or more disease. In one example, theclinical decision support system may have one or more instructionsassociated with taking care of diabetes. By linking the subject'srecords, the clinical decision support system may be able to provideindividualized subject care. For example, by linking the subject'smedical record with the clinical decision support system, the clinicaldecision support system may be able to order additional tests or suggestnext steps based on additional information relating to the subjectincluding but not limited to subject's medical history, subject'sfamily's medical history, demographic information about these subject(age, gender), lifestyle information about the subject (subject's diet,exercise, habits), possible environmental considerations (e.g., if thesubject lived in an area that was exposed to particular toxins or thathas higher risks of certain diseases), and/or any other informationabout the subject.

The clinical decision support system may also be able to providepopulation-based clinical decision support. The clinical decisionsupport system may be able to provide support for one or more peergroups. Such groups may be divided in any manner. For example, thegroups may be based on age, gender, lifestyle, geography, employment,medical history, family medical history, or any other factors. Theclinical decision support system may use epidemiological models forproviding decision support. Information gathered from epidemiologicalsources may be applied to one or more groups of patients.

In one example, an individual may arrive and perform an eligibility testto see if they are eligible for one or more test. The individual maythen be pre-screened and may answer a questionnaire. The questionnairemay include questions about the subject's lifestyle (e.g., diet,exercise, habits) and/or medical history. A physician may perform aphysician check of the individual. In some situations, the questionnaireincludes questions about the subject's dietary consumption, exercise,health condition and/or mental condition. The subject's health conditionmay be of or related to the subject's physiological condition. Thesubject's mental condition may be related to the subject's mood or adepressive disorder, such as depression. The questionnaire may be aguided questionnaire, having a plurality of questions of or related tothe subject's dietary consumption, exercise, health condition and/ormental condition. In some situations, the questionnaire is presented tothe subject with the aid of a system (or sub-system) configured to learnfrom the subject's responses and tailor subsequent questions in responseto the subject's responses. The questionnaire may be presented to thesubject with the aid of a user interface, such as graphical userinterface (GUI), on a display of the device.

In some embodiments, lifestyle recommendations may be made by the deviceand/or system back to the consumer. Such recommendations may be providedprior to, concurrently with, or subsequent to completing thequestionnaire. Such recommendations may be made based on the informationgathered within the questionnaire, medical records, biochemical data,and/or test results.

The device may interpret subject responses to questions with the aid ofreference information. In some situations, the reference informationcomprises a pictorial depiction of portion size of the dietaryconsumption, exertion level of the exercise, existing state of healthcondition and/or existing state of mental condition. The referenceinformation may be included in a calibration matrix stored in a memorylocation (e.g., cache, hard drive, flash memory) of the device.

The device and/or health care personnel may collect biometricinformation about the individual (e.g., blood pressure, weight, bodytemperature). This may be coupled with a test of a sample collected fromthe subject, which may be processed by the device. All the informationmay be linked and may be accessible by the clinical decision supportsystem. In some embodiments, all the information may be linked within asingle subject's records. Such procedures may be useful for annualcheckups or preventative care. Such procedures may also be useful fordiagnosing, treating, or monitoring a disease.

Clinical decision support may provide improved patient triage. Forexample, the clinical decisions support system may make a diagnosis orsuggest a condition of a subject based on the patient's information(e.g., analyte level, physiological information, additional information,or any combination thereof). Such conditions of the patient may bebetter narrowed or more precise/accurate probabilities may be assignedby incorporating the subject-specific information. The clinical decisionsupport may also be able to flag one or more critical situations, andmay cause an alert to be provided to the subject and/or a health careprovider of the subject. The clinical decision support system may beable to flag one or more condition which may require expedited furtheranalysis, and institute one or more proceeding to assist with thefurther analysis.

A health care provider for the subject may be able to access theclinical decision support system and/or additional records associatedwith the subject. For example, the subject may provide a sample to adevice, which may run one or more tests. The clinical decision supportsystem may provide test results to the subject's primary care physician.The primary care physician may be able to view the subject's testresults and/or past test results. The primary care physician may also beable to view additional information provided by the clinical decisionsupport system. In some embodiments, the clinical decision supportsystem may be able to provide the primary care physician withinformation for a specialty outside the primary care physician'sexpertise. For example, if a primary care physician has a cancerpatient, the clinical decision support system may assist the primarycare physician with cancer specific information. The clinical decisionsupport system may provide one or more suggestion to the physician. Thedecision may include one or more recommended intervention by thephysician. Such recommendations may be provided to the physician whenrequested by the physician, when particular conditions are detected,when the clinical decision support is completed with analysis, or upon aschedule. In some embodiments, a device may be provided at thephysician's office. The subject may be able to provide a sample to thedevice at the physician's office, and the physician may receive one ormore test results while the subject is visiting the physician's office.

The clinical decision support system may determine the quality of careof a given health care professional. In some instances, the quality ofcare of a physician may be determined by the clinical decision supportsystem to be provided to one more payer (e.g., health insurancecompany). The quality of care may be determined based on changes in thesubject's data during the subject's interaction with the health careprofessional. Such changes may include lifestyle changes, changes inbiochemical data, feedback from patients, or any other information.

Methods may be provided which may advantageously accommodate reflextesting. Based on one or more test results, additional tests may be runon the device. Such tests and subsequent tests may be scheduled in realtime. Since test results may be provided on-board the device, or may beperformed automatically off-board, and may cause subsequent tests to beautomatically performed using the device. The subsequent tests may beperformed on the same sample upon which one or more initial tests wereperformed. Alternatively, the device may request an additional samplefrom a subject based on the needed tests. After a first test isperformed, if a second test is needed, it may be initiated quickly. Insome embodiments, the second test is initiated in 4 hours or less, 3hours or less, 2 hours or less, 1 hour or less, 30 minutes or less, 15minutes or less, 10 minutes or less, 5 minutes or less, 1 minute orless, 45 seconds or less, 30 seconds or less, 15 seconds or less, 5seconds or less, 1 second or less, or 0.1 second or less from thecompletion of the first test. This may advantageously permit a pluralityof tests to occur without requiring the subject to go to a samplecollection site multiple times. This may also advantageously permit aplurality of tests to occur without requiring a doctor to prescribeadditional steps. The amount of time to reach a diagnosis, monitoring,treatment, and/or prevention of disease may be greatly reduced. Such areflex procedure may be used during a subject's visit to a physician.Such a reflex procedure may occur before the subject sees the physician,while the subject is seeing a physician, and/or after the subject hasseen the physician. The reflex procedure may use the clinical decisionsupport.

In some instances, when a test is ordered, a health care professionalmay do the reflex, and determine additional tests or steps.Alternatively, the device and/or clinical decision support may providereflex testing. For example, if a value is out of range (e.g., level ofan analyte of a sample is outside an expected range), though atouchscreen, a health care professional can do reflex analysis on thesame sample. Alternatively, all tests can be automatically run on asample, and if the health care professional wants to perform anothertest because something is out of range, data can be displayed. In someinstances, the data displayed may only include what the health careprofessional ordered. Alternatively, additional data may be displayedthat may be deemed relevant by the clinical decision support.

In some instances, one or more laboratory report may be provided to ahealth care professional. In some instances, the laboratory report maybe displayed on a sample processing device, or any external device.Laboratory reports and/or laboratory order systems may be customized forreflex analysis. In one example, an order form may permit a user toorder a test, and may also show a field to enter and/or display whatreflex analysis is desired. A report may show reflex analysis that wasconducted for a result. The results of the reflex analysis may also bedisplayed.

The clinical decision support may be capable of self-learning. In someembodiments, a subject's response, a subject's response to one or moretreatment may be monitored, and such data may be accessible by theclinical decision support system. The clinical decision support'sself-learning may be directed to individualized subjects. For example,the clinical decision support may learn that a particular subject doesnot react well to a particular type of drug. The clinical decisionsupport's self-learning may also be generalized. For example, theclinical decision support system may become aware of a pattern thatpeople of a particular demographic or having particular characteristicsmay or may not respond well to a particular treatment. The clinicaldecision support may draw on the subject's records, other patients'records, general health information, public information, medical dataand statistics, insurance information, or other information. Some of theinformation may be publicly available on the Internet (e.g., web sites,articles, journals, databases, medical statistics). The clinicaldecision support system may optionally crawl web sites or databases forupdates to information. Additional information that may becollected/accessed by the clinical decision support system may includean entity's own trials and information about effectiveness and/ortoxicity of drugs. In some embodiments, self-learning may occur on thecloud. As additional data is gathered, it may be uploaded to the cloud,and may be accessible by the clinical decision support system.

The device may be useful for assisting with drug and/or medicationprescriptions. For example, the device may be used to check analytelevels within a subject before a drug prescription is written. Thedevice may determine a drug concentration. The device may be used toperiodically test a subject in order to gauge how much medication thesubject took, regardless of when a re-fill of a medication was made. Thedevice may be used to test a drug presence or level within a subjectprior to, concurrently with, or subsequent to providing a prescriptionfor the drug. Such testing to determine drug levels and/or analytelevels may be useful for testing the efficacy and safety of a drug.After a drug has been prescribed to a subject, the device may be usefulfor determining whether the drug is safe or useful based onpharmacodynamic profiles. Such testing may also be useful for testingthe subject's compliance and/or non-compliance with taking the drug(e.g., if drug levels are too high the subject may be overdosing, ifdrug levels are too low, the subject may not be taking the medication asoften as the subject is supposed to). The device may be useful formonitoring the drug level within the subject over time, to determinewhether the subject is complying with a schedule for taking the drug.Drug and/or analyte levels may be correlated with compliance and/ornon-compliance. A component of the device, such as a blade, may storemedicines, possibly in pill or liquid forms. Based on test results,historical data, physician orders, medical guidance, and/or additionalmedical records as required, such medicines may be dispensed tosubjects. Medicines can be packed, sealed, and labeled as requiredautomatically by the device and then dispensed to the subject.

One or more alert may be provided to a health care professional and/orthe subject if certain conditions are detected. For example, if thedevice is having a toxic or harmful effect on the subject and/or if thesubject is not complying, then appropriate alerts may be provided.

The sample, or a portion thereof, may be archived by the device forlater testing. This process may be triggered by a test result, a deviceerror, or other factors, as defined by a set of procedures and/or rules.The archived sample may be packaged to maintain the integrity of thesample and may be stored in a cooled chamber. The archived sample may besealed in a vessel (e.g. with a septum) and labeled as requiredautomatically by the device. Archived sample may be stored in a vesselunder a vacuum. The archived sample may be later analyzed by the samedevice, or transferred to another device, or sent to another testingfacility. The test results using the archived sample may be combinedwith any prior test results from the initial sample testing.

Devices as described herein may be useful for telemedicine. As describedelsewhere herein, the devices may be useful for verifying the identityof a subject and/or an operator of the device. The device and/or systemmay be able to confirm the subject's identity, access payer information,determine whether the subject received an order to perform a test,determine whether the test falls within a set of rules, access aclinical decision support system, dispense a prescription drug, orperform other steps.

The devices may be capable of performing qualitative and/or quantitativeanalysis of a subject's health and/or medical condition. For example,the devices may be capable processing a sample of the subject, which maybe useful for the determination of one or more analyte level of thesubject. The presence and/or concentration of analyte may be used toassess a health condition of the subject and/or verify the identity ofthe subject. The device may also be capable of collecting one or morephysiological measurement of the subject. Such information may also beuseful to assess the health of the subject and/or verify the identity ofthe subject. In some instances, additional qualitative information aboutthe subject's lifestyle and/or habits may be collected and may be usedto assess the subject's health. Any information collected relating tothe subject as described anywhere herein may be useful for assessing thehealth of the subject (e.g., diagnoses, treatment, and/or diseaseprevention of the subject).

Any information collected by the device relating to the subject may beaccessible by the subject's physician or other health care professional.In some embodiments, only a subset of the information collected by thedevice may be accessible to the subject's physician. Any descriptionherein of a physician may apply to the subject's primary care physician,or other health care professional. The subject's physician may be at aseparate location from the subject. Alternatively, the subject'sphysician may be at the same location from the subject. The subject'sphysician may be able to assess a state of the subject's health withoutseeing the subject in person. The device may be provided at a point ofservice location. The device may advantageously enable a subject to goto a point of service location and have information collected about thesubject which may be relied upon by the physician in assessing thesubject's state of health. The physician may be the subject's primarycare physician, which may enable the subject to maintain personalrelations with a physician that is familiar with the subject, and thesubject's medical history and condition.

In another embodiment, the device or system may perform real timelanguage interpretation services when the patient and the healthcareprovider speak different languages. For instance, a visitor to a countrymay go to a device locations, such as a retail location, connect withthe best medical relevant, qualified or available healthcare person whomay not be able to speak the visitor's language. In that case, thedevice may detect this barrier automatically or the device may promptthe patient or the healthcare provider for language preferences andprovide translation services automatically.

In another embodiment, the device may be placed in a remote andunder-developed area, country or location where large pools ofpopulation may never get access to high quality healthcareprofessionals. In this example, the device, with the help of theexternal controller or the cloud automatically brings healthcare expertsfrom developed world in contact with patients in remote and rural areasand performs language and other cultural interpretations based on notjust spoken language, but sign language, body language and physicalgestures using cameras, image analysis and motion detection and othersensors in the device or modules.

In another embodiment, the device may use the external controller andcloud to overcome certain cultural barriers based on local customs thatprevent delivery of healthcare to certain population. For example, incertain areas where only female healthcare professionals are allowed tointerface with female patients, the device may detect the sex of apatient and automatically or with manual verification connect a femalepatient with a female healthcare provider in a remote or local location,enabling access to greater healthcare services where none or littleaccess to such services would be possible. The device may use imageacquisition, identification, voice and other physical cues using camerasand image analysis and facial recognition to provide this capability.

In some embodiments, the physician may be interacting with the subjectin real time through the device from a remote location, or at the samelocation. In other embodiments, the physician and the subject need notbe interacting in real time—information relating to the subject may becollected via the device, and may be accessible by the physician atanother time. The physician may determine what follow-up actions if anyneed to be made, or whether a real-time in person or remote visit shouldbe scheduled.

One or more camera may be provided which may capture an image of thesubject. Any type of camera or combination of cameras, as describedelsewhere herein may be useful for capturing the image. In someembodiments, the camera may capture a static image of the subject or avideo image of the subject. In one example, a streaming video of asubject may be captured by the device, which may be sent to a physicianat a remote location. A camera may or may not capture an image of thephysician at the physician's location and send and image of thephysician to the device. An image of the physician may be captured by asample processing device at the physician's location. Alternatively, theimage of the physician may be captured by any other type of device. Forexample, the subject and physician may video conference via the device.The video conferencing may show two-dimensional images of the subjectand physician, or three-dimensional images of the subject and physician.In alternate embodiments, audio information may be used forteleconferencing between the subject and physician. One or more staticand/or video images may be captured and sent between the subject and/orphysician.

In some embodiments, conferences may be provided between any number ofparties. For example, a conference may be permitted between two parties(e.g., subject and subject's physician, or subject's primary carephysician and a specialist), three parties (e.g., between the subject,subject's primary care physician, and specialist), four parties, fiveparties, six parties, or more. This may be useful when consulting one ormore specialist or other health care providers for the subject. This mayalso be useful if the subject wishes to loop in a family member orfriend on the conference. Each of the parties may be at separatelocations, or some may be at the same location.

Conversations between the subject and/or physician (or any of theparties or combinations of the parties described herein) may occur inreal-time via the device. Alternatively, the subject may view apre-recorded video of the subject's physician. The subject may record astatement and/or other information from the subject. The recorded videoof the subject may be sent to the subject's physician who may view it inreal-time or at a later time. Any description herein ofsubject-physician interactions may also apply to any other parties,numbers of parties, or combinations described elsewhere herein.

Additionally, images may be captured of a subject, a portion of asubject, or a sample collected from a subject, as described elsewhereherein. Such images may be useful for identification purposes.

Captured images may also be useful for additional purposes. For example,an image may be captured of the subject, and the change or maintenanceof a subject's height and/or girth may be analyzed and assessed forhealth and/or medical purposes. For example, a sudden increase ordecrease in circumference of a subject may raise a red flag or beassessed with other information collected relating to the sample todetermine whether there is a health concern. The subject's gait may beanalyzed to determine if the subject is limping or moving in a way thatindicates an injury. The subject's facial expressions may be stored oranalyzed to determine if the subject is in a particular psychologicalstate.

Images may also be collected of a portion of the body to assess thesubject's state of health. For example, a rash or lesion on thesubject's skin, a mole on the subject's skin, an image of the subject'sthroat, or any other type of image may be collected by the device and/orviewable by the physician. Dermatological conditions may be assessed bythe physician based on one or more image collected of the subject'sskin. Images of one or more of the subject's orifices may be accessibleby the physician. In some embodiments, the images sent may betwo-dimensional images. The images sent may also be three-dimensionalimages, which may be useful in viewing one or more features (e.g.,whether a rash is puffy).

In another example, images of a sample collected from a subject may besent to the physician. For example, one or more images of a tissuesample, bodily fluid sample, or other sample may be sent to thephysician. Images may also include sample at various stages ofprocessing. The device may advantageously be able to produce the imagequickly so that the physician need not wait on such images wheninteracting with the subject. In some embodiments, such images may beaccessible by the subject's primary care physician, pathologist, orother health care professional.

Such images may be analyzed with respect to earlier images collectedwith respect to the subject. Such images may also be analyzed in a standalone fashion without requiring the review of historical imagescollected for the subject. In some embodiments, trend analysis may beperformed on one or more of the images collected from the subject. Suchtrend analysis may extend over a long period of time (e.g., historicaldata relating to a mole on the subject and how it changes over aplurality of visits), or over a shorter period of time (e.g., how asample reacts within the course of a visit). Images from multiple visitsof a subject, or from a single visit of the subject may be analyzed.

In some embodiments, a method for diagnosing or treating a subject withthe aid of the device may be provided. The method may compriseauthenticating a subject and obtaining a three-dimensionalrepresentation of the subject with the aid of a three-dimensionalimaging device. The three-dimensional imaging device may be any of thecameras or plurality of cameras described elsewhere herein. In someembodiments, the three-dimensional imaging device may use a plurality oflenses. The three-dimensional imaging device may include optical, motionand/or audio capture techniques. A system may include an imagerecognition module for analyzing at least a portion of the dynamicthree-dimensional spatial representation of the subject for treatment.The image recognition may or may not be on-board the device. The methodmay include providing the three-dimensional representation to a displayof a computer system of a health care provider, the computer systemcommunicatively coupled to the three-dimensional imaging device, thehealth care provider in remote communication with the subject. Themethod may also include diagnosing or treating the subject with the aidof the three-dimensional representation on the display of the computersystem.

In some instances, the three-dimensional image displayed to thephysician may be an actual three-dimensional image of the portion of thesubject that is imaged. Alternatively, the three-dimensional image maybe representative of the subject captured. This may include simplifiedor modified images. In some embodiments, the three-dimensionalrepresentation may include visual indicators of other informationcollected from the subject. For example, a three dimensional image maybe generated showing a rash on the subject's skin, as well as colorindicators that may be indicative of heat at different areas of therash, or concentrations of analytes detected at different portions ofthe rash. The three-dimensional image may include a computer-generatedmodel.

The health care provider may have been selected by the subject. In someembodiments, the health care provider is the subject's own primary carephysician. The diagnosis may be provided in real-time. In someembodiments, the diagnosis may include combining the three-dimensionalrepresentation with subject specific information. In some embodiments,the subject may be authenticated by verifying the identity of thesubject. Such identification verification may use any of the techniquesdescribed elsewhere herein. In some instances, the subject may beverified via a fingerprint or genetic signature. The subject may beverified by touching a touchscreen of the device. The authenticatingstep may be performed with the aid of one or more of a biometric scan,the subject's insurance card, the subject's name, the subject's driver'slicense, an identification card of the subject, an image of the subjecttaken with the aid of a camera in the point of care system, and agesture-recognition device

A point of service system may be provided for diagnosing or treating asubject. The system may comprise a point of service device having athree-dimensional imaging device for providing a dynamicthree-dimensional spatial representation of the subject; and a remotecomputer system in communication with the three-dimensional imagingdevice, the remote computer system for authenticating the subject and,subsequent to said authenticating, retrieving the dynamicthree-dimensional spatial representation of the subject. The system mayinclude an image recognition module for analyzing at least a portion ofthe dynamic three-dimensional spatial representation of the subject fortreatment.

Other physiological data collected from the subject may be useful forassessing the health of the subject. For example, the subject's bloodpressure level, heart rate, and/or body temperature may be accessed bythe physician and/or may be assessed in view of other informationrelating to the subject to assess the subject's health. The subject'sweight may also be used to assess the subject's health. For example, ifthe subject suddenly gains or loses weight, this may be an indicatorthat may be considered by the physician.

Physical data relating to the subject's sample may be useful forassessing the health of the subject. For example, a sample from thesubject may be processed, and the data collected may be accessible bythe subject's physician. In some embodiments, one or more analyticalsteps may be performed on the data collected by the device before it isviewed by the physician.

Furthermore, as described elsewhere herein, information may be collectedrelating to the subject's lifestyle and/or habits. Such information maybe collected from a graphical user interface, as described elsewhereherein. In some instances, such information may be collected in a surveyform, as described elsewhere herein. In some instances, such informationmay be collected via an external device which may be capable ofcommunicating with the device. The external device may be a computer,server, tablet, mobile device, or any other type of network devicedescribed elsewhere herein. Such information may be stored in the deviceand/or transmitted from the sample processing device. Such informationmay be accessible by a subject's physician or other health careprofessional.

Any information collected relating to the subject may be accessible byone or more physician of the subject, and may be relied upon by thephysician in assessing the health of the subject. Having devices atpoint of service locations may permit a subject to go to one of thepoint of service locations that are convenient to the subject. This maybroaden the subject's access to various physicians. For example, if asubject lives at a first location and has a primary care physician thatthe subject likes, if the subject relocates to a second location, thesubject may still primarily interact with the same primary carephysician. This may also provide flexibility with the subject andphysician's schedules. For example, the subject may provide informationto a sample processing device at a time that the subject is available orwhen convenient for the subject. The physician may be able to accessinformation relating to the subject when the physician has time in thephysician's schedule. In-person and/or real-time meetings or conferencesbetween the physician and subject may be scheduled if/when necessary,but much preliminary data gathering and analysis may occur prior to suchmeetings, thus making such meetings more effective.

Asynchronous Data Management

The systems described herein may optionally use asynchronous datamanagement. Asynchronous data management may use the sample processingdevice described herein. Alternatively, asynchronous data management mayalso occur outside the context of the sample processing device describedherein.

Data may be stored relating to a subject. Such data may include medicalrecords for the subject. Such medical records may span a length of time(e.g., multiple visits), or may be from a single or short point in time(e.g., a single visit). Such data may be accessible by one or moreparties. For example, a subject's physician may be able to access theinformation relating to the subject.

In some embodiments, one or more parties may be able to control who hasaccess to the subject's information, and to which information access isgranted. For example, a subject may determine which physicians or healthcare facilities have access to the subject's data. The subject may wantto choose the subject's physicians and/or specialists. The subject mayspecify which data the other parties have access to. For example, thesubject may determine that certain health care professionals have accessto only a certain subset of medical data. The subject may determine thata specialist only has access to data within the specialist's field orthat may be relevant to the specialist for assessing the health of thesubject. Different parties may be granted access to different subsets ofinformation. Alternatively, the subject may choose to grant differentparties access to the same information. In some instances, the subjectmay choose to grant access to all information.

In some embodiments, other parties may determine who may have access tothe subject's information. For example, a physician's office may collectinformation about the subject. The physician and/or entity affiliatedwith the subject may determine who has access to the information and towhich portions of information the other parties have access to theinformation. In some instances, the physician may determine whichinformation that the subject has access to. In some instances, theinformation collecting entity may determine who has access to which ofthe subject's information. Any other party may be the designated partywho determines who has access to the subject's information.

The granter of access may determine at what time the other parties maybe able to access the selected information. For example, the subject,the physician, or any other party may be the designated granter ofaccess. The granter may provide an expiration time and/or date for theaccess provided to another party. In some instances, the granter mayspecify a start time and/or end time for which the other party canaccess the information. In some instances, the granter need to notspecify an expiration time, and may choose to remove access at any time.

In some instances, the physician may want to share the information withanother health care provider, the subject, or affiliate of the subject.In one example the physician may wish to get a second opinion fromanother health care provider, such as a specialist in a particularfield. The physician may need to get the subject's approval to shareinformation. Alternatively, the physician may have the authority toshare certain portions of information. The first party (e.g., physician)may provide selected data to the second party (e.g., specialist) in afirst format. In one example, the physician may be able to providecharts or other visual depictions of data while including an audioand/or video recording of the physician's thoughts. The data that isshared and/or provided may refer to access that may be granted to theoriginal data.

The second party may view the data in the first format. The second partymay be able to modify the data from the first format to a second format.The second party may be able to insert or modify some of the dataprovided to the second party. For example, the second party may view thecharts or other visual depictions of data with the recording of thephysician's thoughts. The second party may be able to stop the recordingat any point and insert the physician's own thoughts. For example, avideo may be provided showing a visual aspect (e.g., data) and audioaspect (e.g., physician's notes). The second party may be able to stopthe video and record the second party's own voice and thoughts, whichmay be inserted into the video. Similarly, the second party may be ableto modify and manipulate the data shown. For example, the second partymay be able to write the second party's own notes or views into thedisplay of the data.

In addition to adding or inserting additional information, the secondparty may be able to modify the data provided in the first format. Forexample, the first party may draw notes relating to the data. The secondparty may be able to modify the notes—e.g., changing the shape of a lineof a trend, or modifying an equation. The data with the second formatmay be accessible by the second and the first party. In some instances,the second party may send the data in the second party back to the firstparty. Any reference of sending data may include providing access tooriginal data. Original data may be stored in one or more database, orother memory. The original data may be stored in a cloud computing basedinfrastructure.

Such modifications may occur asynchronously. For example, first partymay send information with the first format to the second party. Thesecond party may make such modifications at another time to a secondformat, after the information has been sent. The second party may thensend the information with the second format to the first party. Theinformation may be sent after the modifications have been made. Suchmodifications may manipulate the underlying live data. Discussion ofsending information may relate to sending access to the underlying livedata. In some instances, only one party may access the data to modifythe data at a time. Alternatively, multiple parties may simultaneouslyaccess the data and/or modify the data.

In some embodiments, data may be collected from a sample processingdevice. A sample processing device may also include an interface thatmay permit a user to provide access to one or more other party. Forexample, a send button or interface may be provided where the user canselect the information to send/provide access to, the designatedrecipient(s), and/or time limits. The device may also include a cameraand/or microphone through which the user can record one or more commentsand/or notes that may accompany the data. A user may also be able to addcomments or notes via a touchscreen or other user interface of thedevice.

The data may be stored on the cloud. The user of the device may be ableto select what parties have access to the information. The selectedrecipients may be able to access the data store on the cloud. Theselected recipients may be able to access the data via one or moredevice, which may include a sample processing device, computer, tablet,mobile device, or any other type of network device described elsewhereherein.

In alternate embodiments, such modifications may occur in real-time. Forexample, a video conference may occur where the multiple parties may beviewing the same information at the same time. The conference may permitone or more of the parties to modify the information—e.g., adding notes,drawing figures, or otherwise manipulating the information. The one ormore parties may be manipulating the underlying information, or a visualrepresentation of the information.

Device Calibration and/or Maintenance

In some embodiments the device may be capable of performing on-boardcalibration and/or controls. The device may be capable of performing oneor more diagnostic step (e.g., preparation step and/or assay step). Ifthe results fall outside an expected range, a portion of the device maybe cleaned and/or replaced. The results may also be useful forcalibrating the device. On-board calibration and/or controls may occurwithout requiring human intervention. Calibration and controls may occurwithin a device housing. A device may also be capable of performingon-board maintenance. If during a calibration, operation of device,diagnostic testing, or any other point in time a condition requiringrepair and/or maintenance of the device is detected, the device mayinstitute one or more automated procedures to perform said maintenanceand/or repair. Any description of maintenance may include repair,cleaning, and/or adjustments. For example, a device may detect that acomponent is loose and may automatically tighten the component. Thedevice may also detect that a wash or diluents level is running low in amodule and provide an alert to add more wash or diluents, or bring overwash or diluents from another module.

The system may be configured to continue to function after the removaland/or failure of certain modules.

Calibration and/or maintenance may occur on a periodic basis. In someembodiments, device calibration and/or maintenance may automaticallyoccur at regular or irregular intervals. Device calibration and/ormaintenance may occur when one or more condition is detected from thedevice. For example, if a component appears to be faulty, the device mayrun a diagnostic on associated components. Device calibration and/ormaintenance may occur at the instruction of an operator of the device.Device calibration and/or maintenance may also occur upon automatedinstruction from an external device. The calibration and quality control(QC) cartridge is briefly described in the next paragraph. The goal ofthe calibration cartridge is to enable the quantitative assessment andadjustment of each module/detector of the device. For example, byperforming a variety of assay steps, functionality is tested/evaluatedfor the pipette, gantry, centrifuge, cameras, spectrometer, nucleic acidamplification module, thermal control unit, and cytometer. Eachmeasurement made during calibration cartridge runs with reagent controlsmay be compared to device requirements for precision. By way ofnon-limiting example, there is a pass fail outcome for these results. Ifre-calibration is required, the data generated is used to recalibratethe device (such as the device sensors and pipettes). Recalibrationensures that each device is accurate. Some QC can also be performedautomatically in the device without introducing a cartridge. Forexample, the light sources in the device can be used to periodically QCthe optical sensors in the device. An external device or control maymaintain a device calibration schedule and/or device maintenanceschedule for a plurality of devices. Device calibration and/ormaintenance may occur on a time-based schedule or a use-based schedule.For example, devices that are used more frequently than others may becalibrated and/or maintained more frequently and/or vice versa. QC datamay be indexed with data stored, for example, on the sample processingdevice or an external device.

In some embodiments, a calibration protocol may be stored on a sampleprocessing device, or on an external device and transmitted from theexternal device to the sample processing device. In some embodiments, asample processing device may communicate with an external device toprovide QC data to the external device. In some embodiments, theexternal device may send a protocol or calibration instructions to asample processing device based on QC data provided from the sampleprocessing device to the external device.

In some embodiments, the device may be periodically calibrated andquality controlled. Each module, consisting of one or more hardwareunits, could be calibrated periodically by utilizing a calibrationcartridge. The calibration cartridge may consist of a series of standardfluids, which a properly calibrated system gives a known response to.The module results to these standards could be read, analyzed and basedon deviations or absence thereof, module status can be determined, andcorrected for, if necessary. The calibration standards could either bestored in the device or introduced separately as a cartridge.

In some embodiments, some modules may auto-correct for any changes inthe environment. For example, temperature sensors on the pipette mayautomatically trigger an adjustment in the required piston movement, tocorrect for temperature fluctuations. In general, modules where feedbackregarding performance is available, may auto-correct for any changesover time.

In some embodiments, the output measurements of the cytometer may becalibrated to match results from predicate devices or devices utilizingother technologies as required.

In embodiments, a device may monitor its environment, including itsinternal and external environment. In embodiments, a device may providedevice environmental information to a laboratory. Device environmentalinformation includes, e.g., internal temperature, external temperature,internal humidity, external humidity, time, status of components, errorcodes, images from an internal camera, images from an external camera,and other information. In some embodiments, a device may contain athermal sensor. In embodiments, an internal camera may be fixed at aninternal location. In embodiments, an internal camera may be fixed at aninternal location and may be configured to rotate, scan, or otherwiseprovide views of multiple areas or regions within the device. Inembodiments, an internal camera may be movable within the device; forexample, an internal camera may be mounted on a movable element, such asa pipette, within the device. In embodiments, an internal camera may bemovable within the device and may be configured to rotate, scan, orotherwise provide multiple views of areas within the device frommultiple locations within the device. In embodiments, an external cameramay be fixed at an external location. In embodiments, an external cameramay be fixed at an external location and may be configured to rotate,scan, or otherwise provide multiple views of areas outside the device.In embodiments, an external camera may be movable on or around theoutside of the device. In embodiments, an external camera may be movableand may be configured to rotate, scan, or otherwise provide multipleviews of areas outside the device from multiple locations on or aroundthe outside of the device.

Transmission of device environmental information to a laboratory isuseful for the oversight and control of the device, including beinguseful for the oversight and control of the dynamic operation of thedevice. Transmission of device environmental information to a laboratoryis useful for maintaining the integrity of the operation and control ofthe device, quality control of the operation and control of the device,and for reducing variation or error in the data collection and sampleprocessing performed by the device. For example, transmission oftemperature information to a laboratory is useful for the oversight andcontrol of the device, and is useful in the analysis by the laboratoryof data provided by the device to the laboratory. For example, a devicemay have dedicated temperature zones, and this information may betransmitted to a laboratory.

In embodiments, a device may be configured to control the temperaturewithin the device, or within a portion of the device. The device orportion thereof may be maintained at a single constant temperature, orat a progression of different selected temperatures. Such controlimproves the reproducibility of measurements made within the device, mayunify or provide regularity of conditions for all samples, and reducethe variability of measurements and data, e.g., as measured by thecoefficient of variance of multiple measurements or replicatemeasurements. Such control may also affect chemistry performance in theassay(s) and speed/kinetics of the assay reaction. Temperatureinformation may be useful for quality control. In embodiments, a devicemay monitor temperature and control its internal temperature.Temperature control may be useful for quality control. A device thatmonitors and controls its temperature may transmit temperatureinformation to a laboratory; a laboratory may use such temperatureinformation in the control of the operation of the instrument, in theoversight of the instrument, and in the analysis of data transmittedfrom the instrument. Temperature control may also be used for regulatingthe speed of assays performed with the device. For example, a device maybe maintained at a temperature which optimizes the speed of one or moreselected assays (e.g. at 20° C., 22° C., 25° C., 27° C., 32° C., 35° C.,37° C., 40° C., 42° C., 45° C., 47° C., 50° C., 52° C., 55° C., 57° C.,60° C., 62° C., 65° C., 67° C., 70° C., 72° C., 75° C., 77° C., 80° C.,82° C., 85° C., 87° C., 90° C., 92° C., 95° C., or 97° C.).

In embodiments, a device may be configured to acquire images from withinthe device, or within a portion of the device. Such images may provideinformation about the position, condition, availability, or otherinformation regarding components, reagents, supplies, or samples withinthe device, and may provide information used in control of the operationof the device. Such images may be useful for quality control. A devicethat acquires images from within the device may transmit imageinformation to a laboratory; a laboratory may use such image informationin the control of the operation of the instrument, possibly dynamicallyor in real-time continuously or in real-time but in select intervals, inthe oversight of the instrument, and in the analysis of data transmittedfrom the instrument.

Device Security

One or more security features may be provided on a sample processingdevice. The device may have one or more motion sensor that may determinewhen the device changes orientation or is moved. The device may be ableto detect if someone is trying to open the device. For example one ormore sensor may detect if portions of the device are taken apart. Thedevice may be able to detect if the device falls or is tipped over. Thedevice may be able to sense any motion of the device or any motion nearthe device. For example, the device may be able to sense if an object orperson gets within a certain distance of the device (e.g., using motionsensors, optical sensors, thermal sensors, and/or audio sensors). Thedevice may be able to determine if the device is unplugged or if anerror occurs on the device. Any description of actions that may occur asa result of device tampering may be applied to any other devicecondition as described herein, and vice versa. Accelerometer(s),vibration sensor(s), and/or tilt sensor(s) are used to determine rapidmovements and jarring of the device. Optionally, cameras on the outsideof the device can image and recognize their surroundings and/or providesecurity to the device in terms of video capture, sounding an alert, oronly providing access to verified individual(s) or device(s).

In some embodiments, an alert may be provided if someone is trying toopen a device, or if someone comes within the device's proximity. Insome instances, an alert may be provided if the device housing isbreached. Similarly, an alert may be provided if the device falls, tipsover, or if an error is detected. The device may encompass astabilization system with, optionally, shock absorbance and dampeningcapabilities to prevent it from tipping when for example moving invehicles at high speeds. In some instances, if the device detects thatthe device is being opened, approached, or tampered with, a camera onthe device may capture an image of the device surroundings. The devicemay capture an image of the individual trying to open the device. Thedata associated with the device may be sent to the cloud or an externaldevice. The device associated with the tampering of the device, such asan image of an individual tampering with the device may be transmittedfrom the device. The data associated with the device, which may includeone or more image, may be stored in the device. In the event that thedevice is not able to immediately transmit the data, the data may betransmitted once the device is able and/or connected to a network.

The device may include one or more microphone or audio detection devicethat may be able to record and/or relay sound. For example if a deviceis tampered with, the microphone may collect audio information and theaudio information may be stored on the device or may be transmitted fromthe device.

Optionally, the device may include one or more location sensing device.For example, the device may have a GPS tracker within the device. Whenany tampering with the device is detected, the location of the devicemay be transmitted from the device. The location may be transmitted toan external device or the cloud. In some instances, the location of thedevice may be continuously broadcast once the tampering is detected, ormay be transmitted at one or more intervals or other detected events. Anowner or entity associated with the device may be able to track thelocation of the device. In some instances, a plurality of locationsensors may be provided so that even the device is taken apart and/orone or more location sensor is found and destroyed, it may be possibleto track other parts of the device. In the event that the device isunable to transmit the device location at a particular moment, thedevice may be able to store the device location and transmit it once itis able.

In some embodiments, the device may be designed so that it can only beopened from the inside, or be designed to be only opened from theinside. For example, in some embodiments the device does not havefasteners or screws on the outside of the device. Any mechanicalfastening and/or opening features may be on the inside of the device.The device may be mechanically locked from inside the housing. Theexternal portion of the housing may include no exteriorfastening/locking mechanisms. The device may be opened from the insideupon one or more instructions from a controller. For example, the devicemay have one or more touchscreen or other user interface that may acceptan instruction from a user for the device to open. The device may haveone or more communication unit that may receive an instruction from anexternal device for the device to open. Based on said instructions, oneor more opening mechanism within the device may cause the device toopen. In some instances, the device may require electrical power for thedevice to open. In some instances, the device may only when plugged in.Alternatively, the device may open when powered by a local energystorage system or energy generation system. In some instances, thedevice may only open if it receives instructions from a user who hasbeen identified and/or authenticated. For instance, only certain usersmay be granted the authority to cause the device to open.

The device may have one or more local energy storage system. The energystorage system may permit one or more portions of the device to operateeven if the device is separated from an external energy source. Forexample, if the device is unplugged, one or more energy storage systemmay permit one or more portion of the device to operate. In someinstances, the energy storage system may permit all parts of the deviceto operate. In other examples, the local energy storage system maypermit certain information to be transmitted from the device to thecloud. The local energy storage may be sufficient to power a camera thatmay capture one or more image of the device surroundings and/or anindividual tampering with the device. The local energy storage may besufficient to power a GPS or other location sensor that may indicate thelocation of the device. The local energy storage may be sufficient tosave and/or transmit the state of the device e.g., in a log-basedjournaling approach so that the device can pick up where it left off orknow what steps need to be performed. The local energy storage may besufficient to power a transmission unit that may send informationrelating to the device to the cloud and/or an external device.

In one embodiment, the device and the external controller maintain asecurity mechanism by which no unauthorized person with physical accessto the device may be able to retrieve test information and link it backto an individual, thus protecting the privacy of patient health data. Anexample of this would be where the device captures user identificationinformation, send it to the external device or cloud, receives a secretkey from the cloud and erases all patient information from the device.In such a scenario, if the devices send any further data about thatpatient to the external device, it will be referred to link through thesecret key already obtained from the external device.

Spectrophotometer

Spectrophotometers may contain a light source and an optical sensor, andin some embodiments, may be used for measuring any assay that may bemeasured by assessing an optical property of the assay reaction. Forexample, a spectrophotometer may be used to measure the color,absorbance, transmittance, fluorescence, light-scattering properties, orturbidity of a sample. A spectrophotometer may measure visible light,near-ultraviolet light, or near-infrared light. A spectrophotometer maybe configured to measure a single wavelength of light, or a range ofwavelengths. In some embodiments, a spectrophotometer may measure arange of wavelengths between 100-900 nm, such as, for example 200-600nm, 300-800 nm, 400-800 nm, or 200-800 nm. In some embodiments, aspectrophotometer may measure an optical property of a single sample atmultiple different wavelengths (e.g. the absorbance of a sample atmultiple wavelengths). A spectrophotometer may be configured such thatit may direct light of one or more different wavelengths to a sample andit may detect the transmittance, reflection, or emission of one or moredifferent wavelengths of light by the sample. A spectrophotometer maydirect light of different wavelengths to a sample by, for example, bycontaining contain a monochromator and adjustable filter, such thatlight from the light source may be filtered so that only a selectedwavelength or range of wavelengths reaches the sample. In someembodiments, transmitted light is separated spectrally using a grating,and the spectrally separated signal is read by a spatial sensor. In someembodiments, the light source could be a broad-spectrum light sourcesuch as a Xe, Hg—Xe, Hg—Ar light source. The light source can either bepulsed or continuous, and may allow for adjustable intensity. In anotherexample, a spectrophotometer may contain at least two different lightsources which emit light of different peak wavelength ranges (e.g.different LEDs). A spectrophotometer may also be configured such thatthe optical sensor only detects light of a certain wavelength or rangeof wavelengths (e.g. by use of a filter in front of the sensor). Aspectrophotometer may be a dedicated spectrophotometer (i.e. it may beoptimized for performing spectrophotometric readings; for example, itmay not contain extraneous hardware, such as a sample heater).Optionally, the spectrophotometer may, in certain embodiments, includean electrode or electrochemical detection unit that can be used inconjunction with optical measurements being performed. Optionally, otherhardware such as heating units, cuvette holders, or the like are notexcluded in other embodiments of the spectrophotometer.

FIGS. 74A-74D show a spectrophotometer 7400, in accordance with anembodiment of the invention. The spectrophotometer 7400 may be thespectrophotometer 714 described in the context of FIG. 7. Thespectrophotometer 7400 includes a detection block 7401 (“block”) havinga laser diode, light filter, a sensor (for detecting electromagneticradiation) and a printed circuit board. In some cases, thespectrophotometer 7400 includes a controller having one or moreprocessors. A light source, such as but not limited to a xenon lightsource, is located in a compartment 7402 adjacent the block 7401. Theblock 7401 includes a sample receptacle (or inlet) port or channel 7403,which is configured to accept a first consumable 7404 or a secondconsumable 7405. The first consumable 7404 is a cuvette and the secondconsumable is a tip. The consumables 7404 and 7405 are configured to bemoved, carried and manipulated by various sample handling systems (e.g.,robots) provided herein. The cuvette includes sample holders. Someembodiments may use a light source with specific wavelengths.Optionally, other embodiments do not specifically limit the wavelengths.

With reference to FIG. 74C, the first consumable 7404 is configured tobe mounted in the port 7403. Individual sample holders 7406 of the firstconsumable 7404 are configured to be placed in the line of sight of thelight source 7407 (e.g., xenon light source), either in direct line ofsight or with the aid of optics. Light from the individual sampleholders passes to a detector 7408 (e.g., CCD sensor) for detection. Withreference to FIG. 74D, the second consumable 7405 is inserted into theport 7403 for sample detection. Light from a laser diode 7409 isdirected to the second consumable 7405. Light then passes to a filter7410, which is moved into the path of light emanating from the secondconsumable 7405. Light is then directed to the sensor 7408. Light fromthe first consumable 7404 or second consumable 7405 may be directed tothe sensor 7408 using optics.

The consumables 7404 and 7405 are configured to hold a sample fordetection. The consumables 7404 and 7405 may be discarded after use. Thespectrophotometer 7400 in some cases is configured to hold oneconsumable at a time, though in some situations the spectrophotometer7400 may hold multiple consumables during processing. In somesituations, non-consumable sample holders may be used.

In one embodiment, the fluid handling device might be used to transferan assay vessel into the spectrophotometer where an opticalcharacteristic of the sample is measured. This characteristic mayinclude, but not limited to absorbance, fluorescence, turbidity, etc.The spectrophotometer might include one or more sensors, capable ofhandling one or more sample simultaneously. Analogously, one or moresignals (absorbance, turbidity, etc.) might be measured simultaneously.

The spectrometer may include a PCB board that connects to an externalcomputer and/or processing unit. Alternatively, the computer may be partof the PCB board itself. The computer may receive data from thespectrophotometer sensor, after being processed by the board. Thecomputer may be programmed to analyze the data sent from the board inreal-time. In one embodiment, the results of the computer analysis mayprovide feedback to the board. The feedback may include changes inacquisition time, number of acquisitions for averaging, etc. In someembodiments, this feedback might be used to auto-calibrate thespectrophotometer components.

In some embodiments, the light source and optical sensor of aspectrometer may be oriented in-line with each other. In otherembodiments, the optical sensor is at an angle to the light path fromthe light source (for example, 45 or 90 degrees). An optical sensor atan angle from the light path from the light source may be used, forexample, for detecting light scattered by a sample or light emitted by afluorescent compound.

Referring now to FIG. 74E, yet another embodiment of a spectrophotometerwill now be described. This embodiment shown in FIG. 74E uses adifferent mechanism 7440 for the transport of the cuvette from thecartridge. Instead of using a pipette or other instrument to lift thecuvette out of the pipette and into the detector station, thisembodiment uses a gear in the mechanism 7440 to engage gear teeth 7442formed in the cuvette. This allows for the cuvette 7444 to be moved outfrom the cartridge without having to use a lifting mechanism such as thepipette, a robot, or other end-effector in the system, which then freesthat hardware to perform other tasks. As seen in FIG. 74E, the cuvettemay be moved to detector 7446 which may be single detector or an arrayeddetector.

Referring now to FIGS. 74F and 74G, a still further embodiment of aspectrophotometer will now be described. FIG. 74F is a top-down view ofa fiber-based spectrophotometer wherein the illumination source and/orthe detector can be spaced apart from the sample location and areconnected by fiberoptics 7460 and 7462. This may allow for greaterflexibility in placement of components. Optionally, this also allows forspecific illumination conditions for each sample well of the cuvette,multiple illumination wavelengths per sample well 7464, or other customillumination or detection based on the ability to provide and receivewavelengths of light from and to certain illumination sources anddetectors. By way of non-limiting example, the detector may be a singledetector as shown in this figure or it may be an arrayed detector.

FIG. 74G shows a cutaway perspective view showing the inbound lightpathway 7470 and the outbound light pathway 7472. This embodimentshowing a fiber coupling 7474, a collimator 7476, a mirror 7478, afilter 7480, a reflector 7482, and a fiber coupling 7484 for theoutbound light pathway to the detector. In one embodiment, the samplewell 7464 may be part of a cuvette, or optionally, it may be an openingdesigned to hold a reaction vessel. A fiber-based version of thespectrophotometer can separate the illumination source and the detectorfrom the sample handling unit. The fibers could carry the light sourcefrom a separate location, creating a shared illumination source. Thisprovides for greater flexibility in terms of light source placement andsharing.

With regards to the color strip and the cuvette handling, thespectrophotometer can be configured to accept a single cuvette, multiplecuvettes, or a single cuvette with multiple reaction wells or reactionvessels therein. The positioning of the cuvette by the pipette can be byway of a centralized pipette and the read windows of the cuvette are oneach side of the centralized holder as seen in FIG. K1. Optionally, thepipette can be on or near each end as seen in FIG. K. Optionally, theholder can be on only one end. This can also improve time used forsample preparation if the cuvette has larger number of vessels on thecuvette, particularly if a plurality can be read simultaneously.Optionally, some embodiments may disengage the pipette, robot, or endeffector to drop-off the cuvette at the detector station for detection.During this read or detection time, the sample handling device such asbut not limited to the pipette, robot, or end effector can perform othertask before returning to retrieve the cuvette. It should be understoodthat the cuvette and/or the detector station may have structuralfeatures such as but not limited to lips, edges, legs, or stands thatallow the cuvette to remain upright or in other stable configuration toallow for analyte detection to occur after the drop-off by the samplehandling system.

Optionally, there may be a cuvette that is configured to be disconnectedat the detector and having features such as but not limited to ledges,ridges, lips, hands, or other features to stabilize the cuvette while itis in the detector. The spectrophotometer may have a receiving area thatis shaped to accept this type of cuvette. The system may also beconfigured to accept a single cuvette or have a cuvette that can beloaded with other sample vessels in a sequential, non-simultaneousmanner to provide greater flexibility in scheduling.

Fiberoptics can also provide for multiple channel configurations toenable greater range of excitation and detector configurations. Thefiberoptics can also allow for multiple internal reflections in thecuvette designed to cause this multiple internal reflection path toextend the pathlength beyond the physical geometric pathlength of thecuvette. Some embodiment may have side walls of the cuvette that haveinner wall surfaces with a convex shape such that the vessel that causesreflections of light entering therein.

Assays

Receptor Binding Assays

Receptors:

In some embodiments, the assay station is configured to perform areceptor based assay. In general, receptor based assays comprisedetecting an interaction between two binding partners, an analytereceptor and an analyte. In general, an analyte receptor and an analytein a given pair of binding partners are distinguished on the basis ofwhich one is known (the analyte receptor), and which is being detected(the analyte). As such, exemplary analyte receptors described herein maybe detected as analytes in other embodiments, and exemplary analytes asdescribed herein may be used as analyte receptors for detection ofrespective binding partners in other embodiments. In some embodiments,the analyte receptor, the analyte, or both comprise a protein. Analytereceptors include, but are not limited to: natural or syntheticproteins, cellular receptor proteins, antibodies, enzymes, polypeptides,polynucleotides (e.g. nucleic acid probes, primers, and aptamers),lipids, small organic or inorganic molecules, antigens (e.g. forantibody detection), metal binding ligands, and any other natural orsynthetic molecule having a binding affinity for a target analyte. Insome embodiments, the binding affinity of an analyte receptor for ananalyte is a K_(D) of less than about 5×10⁻⁶M, 1×10⁻⁶M, 5×10⁻⁷M,1×10⁻⁷M, 5×10⁻⁸M, 1×10⁻⁸M, 5×10⁻⁹M, 1×10⁻⁹M, 5×10⁻¹⁰ M, 1×10⁻¹⁰M,5×10⁻¹¹, 1×10⁻¹¹, or less. In some embodiments, an analyte receptordescribed herein (for example, an antibody) may be provided, forexample, in an assay unit, reagent unit, vessel, tip, or container in acartridge or assay station provided herein. Analyte receptors may beprovided in various forms, including, for example, in lyophilized, gel,or liquid forms.

In some embodiments, the analyte receptor is a peptide comprising arecognition structure that binds to a target structure on an analyte,such as a protein. A variety of recognition structures are well known inthe art and can be made using methods known in the art, including byphage display libraries (see, e.g., Gururaja et al. (2000) Chem. Biol.7:515-27; Houimel et al., (2001) Eur. J. Immunol 31:3535-45; Cochran etal. (2001) J. Am. Chem. Soc. 123:625-32; Houimel et al. (2001) Int. J.Cancer 92:748-55, each incorporated herein by reference). A variety ofrecognitions structures are known in the art (see, e.g., Cochran et al.,(2001) J. Am. Chem. Soc. 123:625-32; Boer et al., (2002) Blood100:467-73; Gualillo et al., (2002) Mol. Cell Endocrinol. 190:83-9, eachexpressly incorporated herein by reference), including for examplecombinatorial chemistry methods for producing recognition structuressuch as polymers with affinity for a target structure on a protein (see,e.g., Barn et al., (2001) J. Comb. Chem. 3:534-41; Ju et al., (1999)Biotechnol. 64:232-9, each expressly incorporated herein by reference).

In some embodiments, the analyte receptor is a peptide, polypeptide,oligopeptide or a protein. The peptide, polypeptide, oligopeptide orprotein may be made up of naturally occurring amino acids and peptidebonds, or synthetic peptidomimetic structures. Thus “amino acid”, or“peptide residue”, as used herein include both naturally occurring andsynthetic amino acids. For example, homo-phenylalanine, citrulline andnoreleucine are considered amino acids for the purposes of theinvention. The side chains may be in either the (S) or the (R)configuration. In some embodiments, the amino acids are in the (S) orL-configuration. If non-naturally occurring side chains are used,non-amino acid substituents may be used, for example to prevent orretard in vivo degradation. Proteins comprising non-naturally occurringamino acids may be synthesized or in some cases, made recombinantly;see, for example, van Hest et al., FEBS Lett 428:(1-2) 68-70 May 22,1998 and Tang et al., Abstr. Pap Am. Chem. 5218: U138 Part 2 Aug. 22,1999, both of which are expressly incorporated by reference herein.

In some embodiments, the analyte receptor is cell signaling moleculethat is part of a signaling pathway, such as a receptor protein.Receptor proteins may be membrane associated proteins (e.g.extracellular membrane proteins, intracellular membrane proteins,integral membrane proteins, or transiently membrane-associatedproteins), cytosolic proteins, chaperone proteins, or proteinsassociated with one or more organelles (e.g. nuclear proteins, nuclearenvelope proteins, mitochondrial proteins, golgi and other transportproteins, endosomal proteins, lysosomal proteins, etc.). Examples ofreceptor proteins include, but are not limited to, hormone receptors,steroid receptors, cytokine receptors, such as IL1-α, IL-β, IL-2, IL-3,IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12, IL-15, IL-18, IL-21,CCR5, CCR7, CCR-1-10, CCL20, chemokine receptors, such as CXCR4,adhesion receptors and growth factor receptors, including, but notlimited to, PDGF-R (platelet derived growth factor receptor), EGF-R(epidermal growth factor receptor), VEGF-R (vascular endothelial growthfactor), uPAR (urokinase plasminogen activator receptor), ACHR(acetylcholine receptor), IgE-R (immunoglobulin E receptor), estrogenreceptor, thyroid hormone receptor, CD3 (T cell receptor complex), BCR(B cell receptor complex), CD4, CD28, CD80, CD86, CD54, CD102, CD50,ICAMs (e.g. ICAMs 1, 2 and 3), opioid receptors (mu and kappa), FCreceptors, serotonin receptors (5-HT, 5-HT6, 5-HT7), β-adrenergicreceptors, insulin receptor, leptin receptor, TNF receptor(tissue-necrosis factor), statin receptors, FAS receptor, BAFF receptor,FLT3 LIGAND receptor, GMCSF receptor, and fibronectin receptor. Otherexamples of receptor proteins include the integrin family of receptors.Members of the integrin family of receptors function as heterodimers,composed of various α and β subunits, and mediate interactions between acell's cytoskeleton and the extracellular matrix (reviewed in Giancottiand Ruoslahti, Science 285, 13 Aug. 1999). Different combinations of theα and β subunits give rise to a wide range of ligand specificities,which may be increased further by the presence of cell-type-specificfactors. Integrin clustering is known to activate a number ofintracellular pathways, such as the RAS, Rab, MAP kinase pathway, andthe PI3 kinase pathway. In some embodiments the analyte receptor is aheterodimer composed of a β integrin and an α integrin chosen from thefollowing integrins; β₁, β₂, β₃, β₄, β₅, β₆, α₁, α₂, α₃, α₄, α₅, and α₆,or is MAC-1 (β₂ and cd11b), or αvβ₃. Receptor proteins may be members ofone or more cell signaling pathways, including but not limited to MAPkinase, PI3K/Akt, NFkB, WNT, RAS/RAF/MEK/ERK, JNK/SAPK, p38 MAPK, SrcFamily Kinases, JAK/STAT and/or PKC signaling pathways.

In some embodiments, the analyte receptor is an antibody, and thereceptor-based assay is referred to as an immunoassay having one or moreantigens as analyte. Alternatively, an immunoassay may involve using anantigen as the analyte receptor in order to detect the presence of atarget antibody as an analyte.

In some embodiments, an immunoassay may be an Enzyme-linkedImmunoSorbent assay (“ELISA”). For example, tips having adherentantibodies or target antigens may be used in ELISAs performed bydevices, or on beads in tips/vessels, and according to methods disclosedherein.

Performing an ELISA generally involves at least one antibody capable ofbinding an antigen of interest (i.e., an analyte that is indicative ofinfluenza viral infection). A sample containing or suspected to containthe antigen of interest is immobilized on a support (e.g., a tip orother support having a surface for immobilization) eithernon-specifically (e.g., via adsorption to the surface) or specifically(e.g., via capture by another antibody specific to the same antigen, ina “sandwich” ELISA). After the antigen is immobilized the detectionantibody is added, forming a complex with the antigen. The detectionantibody can be conjugated to an enzyme, or can itself be detected by asecondary antibody which is in turn conjugated to an enzyme. Uponaddition of a substrate for the conjugated enzyme, a detectable signalis generated which indicates the presence and/or quantity of the antigenin the sample. The choice of substrates will depend on the enzymeconjugated. Suitable substrates include fluorogenic and chromogenicsubstrates. One of skill in the art would be knowledgeable as to theparameters that can be modified to increase the signal detected as wellas other variations of ELISAs known in the art.

In some ELISAs, a solid phase capture surface can include an attachedfirst antibody to which a sample (e.g., diluted blood, plasma, orbiological specimen) can be added. If present, an analyte in the samplecan bind to the first antibody and become immobilized. An enzyme reagentcan be added that includes, for example, an antibody coupled orconjugated to an enzyme (e.g., alkaline phosphatase or horseradishperoxidase) that produces a detectable product, or can be otherwisedetected. If the antibody portion of the enzyme reagent can bind theanalyte, then the enzyme reagent also becomes immobilized at the capturesurface. Addition of a substrate for the enzyme can result in a productproducing an effect, for example, light that can be measured andplotted. In this manner the amount of analyte present in a sample can bemeasured.

Thus, for example, an exemplary ELISA which may be performed using adevice, system, or method as disclosed herein includes a solid phasecapture surface (e.g., a tip) on which a first antibody is immobilized.The first antibody is specific for a test antigen (e.g., antibodyspecific for a target blood analyte, such as cholesterol, or for e.g.,neuraminidase on the coat of a virus of interest, or other antigen). Ifthe test antigen is present in a test sample (e.g., whole blood, plasma,or serum) that is exposed to the antibody immobilized on the surface,then the test antigen can become immobilized (captured) at the capturesurface. Addition of a second, labeled antibody that binds to the firstantibody (e.g., where the first antibody is a sheep antibody includingan Fc portion, the second antibody may be an antibody targeting sheep Fcand labeled with alkaline phosphatase) allows the detection andquantification of the amount of antigen in the sample. The firstantibody, which is bound to the substrate, is not washed out by theaddition of the second antibody. Such detection and quantification maybe accomplished, for example, by providing a substrate for the enzymecoupled to the second antibody, leading to the production of colored,fluorescent, luminescent (e.g., chemiluminescent), or otherwisedetectable compounds which may be detected and measured.

Alternatively, after the blood sample is placed in contact with thesurface having the immobilized first antibody (and, optionally, labeledwith an enzyme which catalyzes a reaction that produces a firstdetectable compound) that targets a first antigen, a second antibody,targeting second antigen and labeled with a second enzyme which canproduce a second detectable compound may be added. The first antibody,which is bound to the substrate, is not washed out by the addition ofthe second antibody, and may be detected by providing the substrate andproper reaction conditions for the production of a first detectableproduct by an enzyme linked to the first antibody. Binding andsubsequent detection of the second, labeled antibody at the capturesurface indicates the presence of both the first and the second testantigens in the test sample. Both the first and second detectablecompounds produced by the enzymes linked to the antibodies may bedetected by any means desired, including by detection of fluorescence,luminescence, chemiluminescence, absorbance, colorimetry, or other meansfor detecting the products of the enzymatic reactions due to theattached enzymes.

In some embodiments, photomultipliers tubes, charge-coupled devices,photodiodes, cameras, spectrophotometers, and other components anddevices may be used to measure light emitted or affected during theperformance of an ELISA. For example, the amount of light detected(e.g., in relative light units, or other measurements of luminosity)during the performance of an ELISA on a sample may be compared to astandard curve (e.g., a calibration curve prepared for a particularassay, device, cartridge, or reagent) to calculate the concentration ofthe target analyte in the sample. In some embodiments, any antibodydescribed herein (including antibodies against antigens and pathogensdescribed herein) may be used with an ELISA or optionally in a sandwichimmunoassay.

ELISAs may also be used, for example, in competitive bindingexperiments, in which the concentration of an analyte in a solution maybe measured by adding a known amount of labeled analyte, and measuringthe binding of the analyte. Increased concentrations of the sampleanalyte (which does not include the label) interfere with (“compete”)the binding of the labeled analyte, allowing calculation of the amountof analyte in the sample.

For example, competitive ELISA experiments may be used to determine thebinding characteristics of an antibody or antibody fragment to itstarget. In such experiments, a target analyte is present in solution orbound to a substrate (e.g., a tip, a bead, a microtiter plate).Biotinylated antibodies or antibody fragments may be preincubated withknown concentrations of target in the presence of streptavidin-linkedalkaline phosphatase. After allowing time for incubation, the antibodyor antibody fragment may be allowed to bind to its target, and unboundtarget washed away. Signal may be developed using Alkaline-Phosphatasechemiluminescent substrate and read using a device as disclosed herein,or a separate luminometer, spectrophotometer, or other device.Experimental conditions otherwise identical to those of the testcondition, expect that the solutions do not contain unlabeled target maybe used as baseline measurements as a control. The concentration ofunlabeled target for which 50% of maximal response value is obtained maybe termed the K_(d).

The term “antibody” as used herein refers to immunoglobulin moleculesand immunologically active portions of immunoglobulin molecules, i.e.,molecules that comprise an antigen-binding unit (“Abu” or plural “Abus”)which specifically binds (“immunoreacts with”) an antigen. Structurally,the simplest naturally occurring antibody (e.g., IgG) comprises fourpolypeptide chains, two heavy (H) chains and two light (L) chainsinter-connected by disulfide bonds. The immunoglobulins represent alarge family of molecules that include several types of molecules, suchas IgD, IgG, IgA, IgM and IgE. The term “immunoglobulin molecule”includes, for example, hybrid antibodies, or altered antibodies, andfragments thereof. Antigen-binding unit can be broadly divided into“single-chain” (“Sc”) and “non-single-chain” (“Nsc”) types based ontheir molecular structures.

Also encompassed within the terms “antibodies” and “antigen-bindingunit” are immunoglobulin molecules and fragments thereof that may behuman, nonhuman (vertebrate or invertebrate derived), chimeric, orhumanized. For a description of the concepts of chimeric and humanizedantibodies see Clark et al., 2000 and references cited therein (Clark,(2000) Immunol Today 21:397-402). Chimeric antibodies comprise thevariable region of a nonhuman antibody, for example VH and VL domains ofmouse or rat origin, operably linked to the constant region of a humanantibody (see for example U.S. Pat. No. 4,816,567). In some embodiments,the antibodies of the present invention are humanized. By “humanized”antibody as used herein is meant an antibody comprising a humanframework region (FR) and one or more complementarity determiningregions (CDR's) from a non-human (usually mouse or rat) antibody. Thenon-human antibody providing the CDR's is called the “donor” and thehuman immunoglobulin providing the framework is called the “acceptor”.Humanization relies principally on the grafting of donor CDRs ontoacceptor (human) VL and VH frameworks (Winter U.S. Pat. No. 5,225,539).This strategy is referred to as “CDR grafting”. “Backmutation” ofselected acceptor framework residues to the corresponding donor residuesis often required to regain affinity that is lost in the initial graftedconstruct (U.S. Pat. No. 5,530,101; U.S. Pat. No. 5,585,089; U.S. Pat.No. 5,693,761; U.S. Pat. No. 5,693,762; U.S. Pat. No. 6,180,370; U.S.Pat. No. 5,859,205; U.S. Pat. No. 5,821,337; U.S. Pat. No. 6,054,297;U.S. Pat. No. 6,407,213). The humanized antibody optimally also willcomprise at least a portion of an immunoglobulin constant region,typically that of a human immunoglobulin, and thus will typicallycomprise a human Fc region. Methods for humanizing non-human antibodiesare well known in the art, and can be essentially performed followingthe method of Winter and co-workers (Jones et al., 1986, Nature321:522-525; Riechmann et al., 1988, Nature 332:323-329; Verhoeyen etal., 1988, Science, 239:1534-1536). Additional examples of humanizedmurine monoclonal antibodies are also known in the art, for exampleantibodies binding human protein C (O'Connor et al., 1998, Protein Eng11:321-8), interleukin 2 receptor (Queen et al., 1989, Proc Natl AcadSci, USA 86:10029-33), and human epidermal growth factor receptor 2(Carter et al., 1992, Proc Natl. Acad Sci USA 89:4285-9). In analternate embodiment, the antibodies of the present invention may befully human, that is the sequences of the antibodies are completely orsubstantially human. A number of methods are known in the art forgenerating fully human antibodies, including the use of transgenic mice(Bruggemann et al., 1997, Curr Opin Biotechnol 8:455-458) or humanantibody libraries coupled with selection methods (Griffiths et al.,1998, Curr Opin Biotechnol 9:102-108). Furthermore, the humanizedantibody may comprise residues which are found neither in the recipientantibody nor in the imported CDR or framework sequences. Thesemodifications are made to further refine and optimize antibodyperformance and minimize immunogenicity when introduced into a humanbody.

“Non-single-chain antigen-binding unit” (“Nsc Abus”) are heteromultimerscomprising a light-chain polypeptide and a heavy-chain polypeptide.Examples of the Nsc Abus include but are not limited to (i) a ccFvfragment stabilized by heterodimerization sequences; (ii) any othermonovalent and multivalent molecules comprising at least one ccFvfragment; (iii) an Fab fragment consisting of the VL, VH, CL and CH1domains; (iv) an Fd fragment consisting of the VH and CH1 domains; (v)an Fv fragment consisting of the VL and VH domains of a single arm of anantibody; (vi) an F(ab′)2 fragment, a bivalent fragment comprising twoFab fragments linked by a disulfide bridge at the hinge region; (vii) adiabody; and (viii) any other Nsc Abus that are described in Little etal. (2000) Immunology Today, or in U.S. Pat. No. 7,429,652.

As noted above, a Nsc Abus can be either “monovalent” or “multivalent.”Whereas the former has one binding site per antigen-binding unit, thelatter contains multiple binding sites capable of binding to more thanone antigen of the same or different kind. Depending on the number ofbinding sites, a Nsc Abus may be bivalent (having two antigen-bindingsites), trivalent (having three antigen-binding sites), tetravalent(having four antigen-binding sites), and so on.

Multivalent Nsc Abus can be further classified on the basis of theirbinding specificities. A “monospecific” Nsc Abu is a molecule capable ofbinding to one or more antigens of the same kind. A “multispecific” NscAbu is a molecule having binding specificities for at least twodifferent antigens. While such molecules normally will only bind twodistinct antigens (i.e. bispecific Abus), antibodies with additionalspecificities such as trispecific antibodies are encompassed by thisexpression when used herein. Examples of bispecific antigen bindingunits include those with one arm directed against a tumor cell antigenand the other arm directed against a cytotoxic trigger molecule such asanti-CD3/anti-malignant B-cell (1D10), anti-CD3/anti-p185 HER2,anti-CD3/anti-p97, anti-CD3/anti-renal cell carcinoma,anti-CD3/anti-OVCAR-3, anti-CD3/L-D1 (anti-colon carcinoma),anti-CD3/anti-melanocyte stimulating hormone analog,anti-FcγRI/anti-CD15, anti-p185 HER2/FcγRIII (CD16), anti-EGFreceptor/anti-CD3, anti-CD3/anti-CAMA1, anti-CD3/anti-CD19, anti-CD3/MoV18, anti-FcγR/anti-HIV; bispecific Abus for tumor detection in vitro orin vivo such as anti-CEA/anti-EOTUBE, anti-CEA/anti-DPTA, anti-p 185HER2/anti-hapten; BsAbs as vaccine adjuvants (see Fanger et al., supra);and bispecific Abus as diagnostic tools such as anti-rabbitIgG/anti-ferritin, anti-horse radish peroxidase (HRP)/anti-hormone,anti-somatostatin/anti-substance P, anti-neural cell ahesion molecule(NCAM)/anti-CD3, anti-folate binding protein (FBP)/anti-CD3, anti-pancarcinoma associated antigen (AMOC-31)/anti-CD3; bispecific Abus withone arm which binds specifically to a tumor antigen and one arm whichbinds to a toxin such as anti-saporin/anti-Id-1, anti-CD22/anti-saporin,anti-CD7/anti-saporin, anti-CD38/anti-saporin, anti-CEA/anti-ricin Achain, anti-interferon-α (IFN-α)/anti-hybridoma idiotype,anti-CEA/anti-vinca alkaloid; BsAbs for converting enzyme activatedprodrugs such as anti-CD30/anti-alkaline phosphatase (which catalyzesconversion of mitomycin phosphate prodrug to mitomycin alcohol);bispecific Abus which can be used as fibrinolytic agents such asanti-fibrin/anti-tissue plasminogen activator (tPA),anti-fibrin/anti-urokinase-type plasminogen activator (uPA); bispecificantigen-binding units for targeting immune complexes to cell surfacereceptors such as anti-low density lipoprotein (LDL)/anti-Fc receptor(e.g. Fcγ RI, FcγRII or FcγRIII); bispecific Abus for use in therapy ofinfectious diseases such as anti-CD3/anti-herpes simplex virus (HSV),anti-T-cell receptor:CD3 complex/anti-influenza, anti-HRP/anti-FITC,anti-CEA/anti-β-galactosidase (see Nolan et al., supra). Examples oftrispecific antibodies include anti-CD3/anti-CD4/anti-CD37,anti-CD3/anti-CD5/anti-CD37 and anti-CD3/anti-CD8/anti-CD37.

“Single-chain antigen-binding unit” (“Sc Abu”) refers to a monomericAbu. Although the two domains of the Fv fragment are coded for byseparate genes, a synthetic linker can be made that enables them to bemade as a single protein chain (i.e. single chain Fv (“scFv”) asdescribed in Bird et al. (1998) Science 242:423-426 and Huston et al.1988) PNAS 85:5879-5883) BY RECOMBINANT METHODS. Other Sc Abus includeantigen-binding molecules stabilized by heterodimerization sequences,and dAb fragments (Ward et al., (1989) Nature 341:544-546) which consistof a VH domain and an isolated complementarity determining region (CDR).An example of a linking peptide is a sequence of four glycines followedby a serine, the sequence of 5 amino acids repeated twice for a totallength of 15 amino acids, which linking peptide bridges approximately3.5 nm between the carboxyl terminus of one V region and the aminoterminus of another V region. Other linker sequences can also be used,and can provide additional functions, such as a means for attaching adrug or a solid support. A preferred single-chain antigen-binding unitcontains VL and VH regions that are linked together and stabilized by apair of subject heterodimerization sequences. The scFvs can be assembledin any order, for example, VH-(first heterodimerizationsequence)-(second heterodimerization sequence)-VL, or VL-(firstheterodimerization sequence)-(second heterodimerization sequence)-VH. Anantibody or Abu “specifically binds to” or “immunoreactive with” anantigen if it binds with greater affinity or avidity than it binds toother reference antigens including polypeptides or other substances.

In some embodiments, the analyte receptor is an enzyme and the targetanalyte is a substrate of the enzyme, or the analyte receptor is anenzyme substrate and the analyte is an enzyme that acts on thesubstrate, such that detection is effected by the activity of the enzymeon the substrate, such as by the production of a detectable product.Many enzymes useful in the detection of or detectable by activity onvarious substrates are known in the art, and include without limitation,proteases, phosphatases, peroxidases, sulfatases, peptidases,glycosidases, hydrolases, oxidoreductases, lyases, transferases,isomerases, ligases, and synthases, Of particular interest are classesof enzymes that have physiological significance. These enzymes include,without limitation, protein kinases, peptidases, esterases, proteinphosphatases, isomerases, glycosylases, synthetases, proteases,dehydrogenases, oxidases, reductases, methylases and the like. Enzymesof interest include those involved in making or hydrolyzing esters, bothorganic and inorganic, glycosylating, and hydrolyzing amides. In anyclass, there may be further subdivisions, as in the kinases, where thekinase may be specific for phosphorylation of serine, threonine and/ortyrosine residues in peptides and proteins. Thus, the enzymes may be,for example, kinases from different functional groups of kinases,including cyclic nucleotide-regulated protein kinases, protein kinase C,kinases regulated by Ca.sup.2+/CaM, cyclin-dependent kinases, ERK/MAPkinases, and protein-tyrosine kinases. The kinase may be a proteinkinase enzyme in a signaling pathway, effective to phosphorylate anoligopeptide substrate, such as ERK kinase, S6 kinase, IR kinase, P38kinase, and AbI kinase. For these, the substrates can include anoligopeptide substrate. Other kinases of interest may include, forexample, Src kinase, JNK, MAP kinase, cyclin-dependent kinases, P53kinases, platelet-derived growth factor receptor, epidermal growthfactor receptor, and MEK.

In particular, enzymes that are useful in the present invention includeany protein that exhibits enzymatic activity, e.g., lipases,phospholipases, sulphatases, ureases, peptidases, proteases andesterases, including acid phosphatases, glucosidases, glucuronidases,galactosidases, carboxylesterases, and luciferases. In one embodiment,one of the enzymes is a hydrolytic enzyme. In another embodiment, atleast two of the enzymes are hydrolytic enzymes. Examples of hydrolyticenzymes include alkaline and acid phosphatases, esterases,decarboxylases, phospholipase D, P-xylosidase, β-D-fucosidase,thioglucosidase, β-D-galactosidase, α-D-galactosidase, α-D-glucosidase,β-D-glucosidase, β-D-glucuronidase, α-D-mannosidase, β-D-mannosidase,β-D-fructofuranosidase, and β-D-glucosiduronase. In some embodiments,the product of the enzyme directly produces a detectable feature in areaction (e.g. change in color, turbidity, absorbance of a wavelength oflight, fluorescence, chemiluminescence, electrical conductance, ortemperature). In some embodiments, the product of the enzyme is detectedindirectly by binding of a second analyte receptor having a detectablelabel.

In some embodiments, an analyte receptor used to detect an analyte is anaptamer. An aptamer can be on a bead or other surface, such as a microarray-type surface. The term “aptamer” is used to refer to a peptide,nucleic acid, or a combination thereof that is selected for the abilityto specifically bind one or more target analytes. Peptide aptamers areaffinity agents that generally comprise one or more variable loopdomains displayed on the surface of a scaffold protein. A nucleic acidaptamer is a specific binding oligonucleotide, which is anoligonucleotide that is capable of selectively forming a complex with anintended target analyte. The complexation is target-specific in thesense that other materials, such as other analytes that may accompanythe target analyte, do not complex to the aptamer with as great anaffinity. It is recognized that complexation and affinity are a matterof degree; however, in this context, “target-specific” means that theaptamer binds to target with a much higher degree of affinity than itbinds to contaminating materials. The meaning of specificity in thiscontext is thus similar to the meaning of specificity as applied toantibodies, for example. The aptamer may be prepared by any knownmethod, including synthetic, recombinant, and purification methods.Further, the term “aptamer” also includes “secondary aptamers”containing a consensus sequence derived from comparing two or more knownaptamers to a given target.

In general, nucleic acid aptamers are about 9 to about 35 nucleotides inlength. In some embodiments, a nucleic acid aptamer is at least 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65,70, 80, 90, 100, or more nucleic acids in length. Although theoligonucleotides of the aptamers generally are single-stranded ordouble-stranded, it is contemplated that aptamers may sometimes assumetriple-stranded or quadruple-stranded structures. In some embodiments, anucleic acid aptamer is circular, such as in US20050176940. The specificbinding oligonucleotides of the aptamers should contain thesequence-conferring specificity, but may be extended with flankingregions and otherwise derivatized or modified. The aptamers found tobind to a target analyte may be isolated, sequenced, and thenre-synthesized as conventional DNA or RNA moieties, or may be modifiedoligomers. These modifications include, but are not limited toincorporation of: (1) modified or analogous forms of sugars (e.g. riboseand deoxyribose); (2) alternative linking groups; or (3) analogous formsof purine and pyrimidine bases.

Nucleic acid aptamers can comprise DNA, RNA, functionalized or modifiednucleic acid bases, nucleic acid analogues, modified or alternativebackbone chemistries, or combinations thereof. The oligonucleotides ofthe aptamers may contain the conventional bases adenine, guanine,cytosine, and thymine or uridine. Included within the term aptamers aresynthetic aptamers that incorporate analogous forms of purines andpyrimidines. “Analogous” forms of purines and pyrimidines are thosegenerally known in the art, many of which are used as chemotherapeuticagents. Non-limiting examples of analogous forms of purines andpyrimidines (i.e. base analogues) include aziridinylcytosine,4-acetylcytosine, 5-fluorouracil, 5-bromouracil,5-carboxymethylaminomethyl-2-thiouracil,5-carboxymethyl-aminomethyluracil, inosine, N6-isopentenyladenine,1-methyladenine, 1-methylpseudouracil, 1-methylguanine, 1-methylinosine,2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine,5-methylcytosine, N6-methyladenine, 7-methylguanine,5-methylaminomethyl-uracil, 5-methoxyaminomethyl-2-thiouracil,beta-D-mannosylqueosine, 5-methoxyuracil,2-methyl-thio-N6-isopentenyladenine, uracil-5-oxyacetic acidmethylester, pseudouracil, queosine, 2-thiocytosine,5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil,uracil-5-oxyacetic acid, 5-pentynyl-uracil, and 2,6-diaminopurine. Theuse of uracil as a substitute base for thymine in deoxyribonucleic acid(hereinafter referred to as “dU”) is considered to be an “analogous”form of pyrimidine in this invention.

Aptamer oligonucleotides may contain analogous forms of ribose ordeoxyribose sugars that are known in the art, including but not limitedto 2′ substituted sugars such as 2′-O-methyl-, 2′-O-allyl, 2′-fluoro- or2′-azido-ribose, carbocyclic sugar analogs, alpha-anomeric sugars,epimeric sugars such as arabinose, xyloses or lyxoses, pyranose sugars,furanose sugars, sedoheptuloses, locked nucleic acids (LNA), peptidenucleic acid (PNA), acyclic analogs and abasic nucleoside analogs suchas methyl riboside.

Aptamers may also include intermediates in their synthesis. For example,any of the hydroxyl groups ordinarily present may be replaced byphosphonate groups, phosphate groups, protected by a standard protectinggroup, or activated to prepare additional linkages to additionalnucleotides or substrates. The 5′ terminal OH is conventionally free butmay be phosphorylated; OH substituents at the 3′ terminus may also bephosphorylated. The hydroxyls may also be derivatized to standardprotecting groups. One or more phosphodiester linkages may be replacedby alternative linking groups. These alternative linking groups include,but are not limited to embodiments wherein P(O)O is replaced by P(O)S(“thioate”), P(S)S (“dithioate”), P(O)NR 2 (“amidate”), P(O)R, P(O)OR′,CO or CH 2 (“formacetal”), wherein each R or R′ is independently H orsubstituted or unsubstituted alkyl (1-20C.) optionally containing anether (—O—) linkage, aryl, alkenyl, cycloalkyl, cycloalkenyl or aralkyl.

One particular embodiment of aptamers that are useful in the presentinvention is based on RNA aptamers as disclosed in U.S. Pat. Nos.5,270,163 and 5,475,096, which are incorporated herein by reference. Theaforementioned patents disclose the SELEX method, which involvesselection from a mixture of candidate oligonucleotides and stepwiseiterations of binding, partitioning and amplification, using the samegeneral selection scheme, to achieve virtually any desired criterion ofbinding affinity and selectivity. Starting from a mixture of nucleicacids, preferably comprising a segment of randomized sequence, the SELEXmethod includes steps of contacting the mixture with a target, such as atarget analyte, under conditions favorable for binding, partitioningunbound nucleic acids from those nucleic acids which have boundspecifically to target molecules, dissociating the nucleic acid-targetcomplexes, amplifying the nucleic acids dissociated from the nucleicacid-target complexes to yield a ligand-enriched mixture of nucleicacids, then reiterating the steps of binding, partitioning, dissociatingand amplifying through as many cycles as desired to yield highlyspecific, high affinity nucleic acid ligands to the target molecule. Insome embodiments, negative screening is employed in which a plurality ofaptamers are exposed to analytes or other materials likely to be foundtogether with target analytes in a sample to be analyzed, and onlyaptamers that do not bind are retained.

The SELEX method encompasses the identification of high-affinity nucleicacid ligands containing modified nucleotides conferring improvedcharacteristics on the ligand, such as improved in vivo stability orimproved delivery characteristics. Examples of such modificationsinclude chemical substitutions at the ribose and/or phosphate and/orbase positions. In some embodiments, two or more aptamers are joined toform a single, multivalent aptamer molecule. Multivalent aptamermolecules can comprise multiple copies of an aptamer, each copytargeting the same analyte, two or more different aptamers targetingdifferent analytes, or combinations of these.

Analyte receptors can be used to detect an analyte in any of thedetection schemes described herein. In one embodiment, analyte receptorsare covalently or non-covalently coupled to a substrate. Non-limitingexamples of substrates to which analyte receptors may be coupled includemicroarrays, microbeads, pipette tips, sample transfer devices,cuvettes, capillaries or other tubes, reaction chambers, or any othersuitable format compatible with the subject detection system. Biochipmicroarray production can employ various semiconductor fabricationtechniques, such as solid phase chemistry, combinatorial chemistry,molecular biology, and robotics. One process typically used is aphotolithographic manufacturing process for producing microarrays withmillions of analyte receptors on a single chip. Alternatively, if theanalyte receptors are pre-synthesized, they can be attached to an arraysurface using techniques such as micro-channel pumping,“ink-jet”spotting, template-stamping, or photocrosslinking. An exemplaryphotolithographic process begins by coating a quartz wafer with alight-sensitive chemical compound to prevent coupling between the quartzwafer and the first nucleotide of a DNA probe being created. Alithographic mask is used to either inhibit or permit the transmissionof light onto specific locations of the wafer surface. The surface isthen contacted with a solution which may contain adenine, thymine,cytosine, or guanine, and coupling occurs only in those regions on theglass that have been deprotected through illumination. The couplednucleotide bears a light-sensitive protecting group, allowing the cyclecan be repeated. In this manner, the microarray is created as the probesare synthesized via repeated cycles of deprotection and coupling. Theprocess may be repeated until the probes reach their full length.Commercially available arrays are typically manufactured at a density ofover 1.3 million unique features per array. Depending on the demands ofthe experiment and the number of probes required per array, each wafer,can be cut into tens or hundreds of individual arrays.

Other methods may be used to produce a coated solid surface with analytereceptors attached thereto. A coated solid surface may be aLangmuir-Bodgett film, functionalized glass, germanium, silicon, PTFE,polystyrene, gallium arsenide, gold, silver, membrane, nylon, PVP,polymer plastics, or any other material known in the art that is capableof having functional groups such as amino, carboxyl, Diels-Alderreactants, thiol or hydroxyl incorporated on its surface. These groupsmay then be covalently attached to crosslinking agents, so that thesubsequent binding of the analyte receptors and target analyte willoccur in solution without hindrance from the biochip. Typicalcrosslinking groups include ethylene glycol oligomer, diamines, andamino acids. Alternatively, analyte receptors may be coupled to an arrayusing enzymatic procedures, such as described in US20100240544.

In some embodiments, analyte receptors are coupled to the surface of amicrobead. Microbeads useful in coupling to analyte receptors, such asoligonucleotides, are known in the art, and include magnetic andnon-magnetic beads. Microbeads can be labeled with 1, 2, 3, 4, 5, 6, 7,8, 9, 10, or more dyes to facilitate coding of the beads andidentification of an analyte receptor joined thereto. Coding ofmicrobeads can be used to distinguish at least 10, 50, 100, 200, 300,400, 500, 600, 700, 800, 900, 1000, 1500, 2000, 5000, or more differentmicrobeads in a single assay, each microbead corresponding to adifferent analyte receptors with specificity for a different analyte.

In some embodiments, analyte receptors are coupled to the surface of areaction chamber, such as a tip. For example, the interior surface of atip may be coated with an analyte receptor specific for a singleanalyte. Alternatively, the interior surface of a tip may be coated withtwo or more different analyte receptors specific for different analytes.When two or more different analyte receptors are coupled to the sameinterior tip surface, each of the different analyte receptors may becoupled at different known locations, such as forming distinct orderedrings or bands at different positions along the axis of a tip. In thiscase, multiple different analytes may be analyzed in the same sample bydrawing a sample up a tip and allowing analytes contained in the sampleto bind with the analyte receptors coated at successive positions alongthe tip. Binding events can then be visualized as described herein, withthe location of each band in a banding pattern corresponding to aspecific known analyte.

Analytes:

Analyte receptors can be used as diagnostic and prognostic reagents, asreagents for the discovery of novel therapeutics, as reagents formonitoring drug response in individuals, and as reagents for thediscovery of novel therapeutic targets. Analyte receptors can be used todetect one or more target analytes. The term “analytes” refers to anytype of biological molecule including, for example, simple intermediarymetabolites, sugars, lipids, and hormones as well as macromolecules suchas complex carbohydrates, phospholipids, nucleic acids (e.g. DNA, RNA,mRNA, miRNA, rRNA, tRNA), polypeptides and peptides. Furthernon-limiting examples of analytes include drugs, drug candidates,prodrugs, pharmaceutical agents, drug metabolites, biomarkers such asexpressed proteins and cell markers, antibodies, serum proteins,cholesterol and other metabolites, electrolytes, metal ions,polysaccharides, genes, proteins, glycoproteins, glycolipids, lectins,growth factors, cytokines, vitamins, enzymes, enzyme substrates, enzymeinhibitors, steroids, oxygen and other gases found in physiologic fluids(e.g. CO₂), cells, cellular constituents, cell adhesion molecules, plantand animal products, cell surface markers (e.g. cell surface receptorsand other molecules identified herein as receptor proteins), and cellsignaling molecules. Non-limiting examples of protein analytes includemembrane associated proteins (e.g. extracellular membrane proteins,intracellular membrane proteins, integral membrane proteins, ortransiently membrane-associated proteins), cytosolic proteins, chaperoneproteins, proteins associated with one or more organelles (e.g. nuclearproteins, nuclear envelope proteins, mitochondrial proteins, golgi andother transport proteins, endosomal proteins, lysosomal proteins, etc.),secreted proteins, serum proteins, and toxins. Non-limiting examples ofanalytes for detection include Adiponectin, Alanine Aminotransferase(ALT/GPT), Alpha-fetoprotein (AFP), Albumin, Alkaline Phosphatase (ALP),Alpha Fetoprotein, Apolipoprotein A-I (Apo A-I), Apolipoprotein B (ApoB), Apolipoprotein B/Apoplipoprotien A-1 Ratio (Apo B/A1 ratio),Aspartate Aminotransferase (AST/GOT), AspirinWorks®(11-Dehydro-Thromboxane B2), Bicarbonate (CO2), Bilirubin, Direct(DBIL), Bilirubin, Total (TBIL), Blood Urea Nitrogen (BUN), Carboxyterminal collagen crosslinks (Beta-CrossLaps), Calcium, Cancer Antigen125 (CA 125), Cancer Antigen 15-3 (CA 15-3), Cancer Antigen 19-9 (CA19-9), Carcinoembryonic Antigen (CEA), Chloride (Cl), Complete BloodCount w/differential (CBC), C-peptide, C-reactive protein (CRP-hs),Creatine Kinase (CK), Creatinine (serum), Creatinine (urine), CytochromeP450, Cystatin-C, D-Dimer, Dehydroepiandrosterone Sulfate (DHEA-S),Estradiol, F2 Isoprostanes, Factor V Leiden, Ferritin, Fibrinogen(mass), Folate, Follicle-stimulating Hormone (FSH), Free FattyAcids/Non-Esterified Fatty Acids (FFA/NEFA), Fructosamine,Gamma-glutamyl Transferase (GGT), Glucose, HbA1c & estimated AverageGlucose (eAG), HDL2 subclass, High-density Lipoprotein Cholesterol(HDL-C), High-density Lipoprotein Particle Number (HDL-P),High-sensitivity C-reactive Protein (hs-CRP), Homocysteine, Insulin,Iron and TIBC, Lactate dehydrogenase (LDH), Leptin, Lipoprotein (a)Cholesterol (Lp(a) chol), Lipoprotein (a) Mass (Lp(a) mass),Lipoprotein-associated Phospholipase A2 (Lp-PLA2), Low-densityLipoprotein Cholesterol, Direct (LDL-C), Low-density LipoproteinParticle Number (LDL-P), Luteinizing Hormone (LH), Magnesium,Methylenetetrahydrofolate reductase (MTHFR), Micro-albumin,Myeloperoxidase (MPO), N-terminal Pro b-type Natriuretic Peptide(NT-proBNP), Non-High-density Lipoprotein Cholesterol, Omega-3 FattyAcid Profile, Osteocalcin, Parathyroid Hormone (PTH), Phosphorus,Potassium (K+), Prostate Specific Antigen, total (PSA, total),Prothrombin, Resistin, Sex Hormone Binding Globulin (SHBG), Small DenseLow-density Lipoprotein (sdLDL), Small dense low-densityLipoprotein/Low-density Lipoprotein Cholesterol Ratio (sd LDL/LDL-Cratio), Sodium (NA+), T Uptake, Testosterone, Thyroid-stimulatinghormone (TSH), Thyroxine (T4), Total Cholesterol (TCHOL), Total Protein,Triglycerides (TRIG), Triiodothyronine (T3), T4 (free), Uric Acid,Vitamin B12, 25-hydroxy-vitamin D, clotting factors (e.g. factor I(fibrinogen), factor II (prothrombin), factor III (tissuethromboplastin), factor IV (calcium), factor V (proaccelerin), factor VI(no longer considered active in hemostasis), factor VII (proconvertin),factor VIII (antihemophilic factor), factor IX (plasma thromboplastincomponent; Christmas factor), factor X (stuart factor), factor XI(plasma thromboplastin antecedent), factor XII (hageman factor), factorXIII (fibrin stabilizing factor)).

In some embodiments, the analyte is a cell signaling molecule, such as aprotein. Non-limiting examples of proteins that may be detected asanalytes include kinases, phosphatases, lipid signaling molecules,adaptor/scaffold proteins, GTPase activating proteins, isomerases,deacetylases, methylases, demethylases, tumor suppressor genes,caspases, proteins involved in apoptosis, cell cycle regulators,molecular chaperones, metabolic enzymes, vesicular transport proteins,cytokines, cytokine regulators, ubiquitination enzymes, adhesionmolecules, cytoskeletal/contractile proteins, heterotrimeric G proteins,small molecular weight GTPases, guanine nucleotide exchange factors,hydroxylases, proteases, ion channels, molecular transporters,transcription factors/DNA binding factors, regulators of transcription,and regulators of translation. Analytes may be members of any cellsignaling pathway, including but not limited to MAP kinase, PI3K/Akt,NFkB, WNT, RAS/RAF/MEK/ERK, JNK/SAPK, p38 MAPK, Src Family Kinases,JAK/STAT and/or PKC signaling pathways. Examples of signaling moleculesinclude, but are not limited to, HER receptors, PDGF receptors, Kitreceptor, FGF receptors, Eph receptors, Trk receptors, IGF receptors,Insulin receptor, Met receptor, Ret, VEGF receptors, TIE1, TIE2, FAK,Jak1, Jak2, Jak3, Tyk2, Src, Lyn, Fyn, Lck, Fgr, Yes, Csk, Abl, Btk,ZAP70, Syk, IRAKs, cRaf, ARaf, BRAF, Mos, Lim kinase, ILK, Tpl, ALK,TGFβ receptors, BMP receptors, MEKKs, ASK, MLKs, DLK, PAKs, Mek 1, Mek2, MKK3/6, MKK4/7, ASK1, Cot, NIK, Bub, Myt 1, Wee1, Casein kinases,PDK1, SGK1, SGK2, SGK3, Akt1, Akt2, Akt3, p90Rsks, p70S6 Kinase, Prks,PKCs, PKAs, ROCK 1, ROCK 2, Auroras, CaMKs, MNKs, AMPKs, MELK, MARKs,Chk1, Chk2, LKB-1, MAPKAPKs, Pim1, Pim2, Pim3, IKKs, Cdks, Jnks, Erks,IKKs, GSK3a, GSK3β, Cdks, CLKs, PKR, PI3-Kinase class 1, class 2, class3, mTor, SAPK/JNK1,2,3, p38s, PKR, DNA-PK, ATM, ATR, Receptor proteintyrosine phosphatases (RPTPs), LAR phosphatase, CD45, Non receptortyrosine phosphatases (NPRTPs), SHPs, MAP kinase phosphatases (MKPs),Dual Specificity phosphatases (DUSPs), CDC25 phosphatases, Low molecularweight tyrosine phosphatase, Eyes absent (EYA) tyrosine phosphatases,Slingshot phosphatases (SSH), serine phosphatases, PP2A, PP2B, PP2C,PP1, PPS, inositol phosphatases, PTEN, SHIPs, myotubularins,phosphoinositide kinases, phopsholipases, prostaglandin synthases,5-lipoxygenase, sphingosine kinases, sphingomyelinases, adaptor/scaffoldproteins, Shc, Grb2, BLNK, LAT, B cell adaptor for PI3-kinase (BCAP),SLAP, Dok, KSR, MyD88, Crk, CrkL, GAD, Nck, Grb2 associated binder(GAB), Fas associated death domain (FADD), TRADD, TRAF2, RIP, T-Cellleukemia family, IL-2, IL-4, IL-8, IL-6, interferon β, interferon α,suppressors of cytokine signaling (SOCs), Cbl, SCF ubiquitination ligasecomplex, APC/C, adhesion molecules, integrins, Immunoglobulin-likeadhesion molecules, selectins, cadherins, catenins, focal adhesionkinase, p130CAS, fodrin, actin, paxillin, myosin, myosin bindingproteins, tubulin, eg5/KSP, CENPs, β-adrenergic receptors, muscarinicreceptors, adenylyl cyclase receptors, small molecular weight GTPases,H-Ras, K-Ras, N-Ras, Ran, Rac, Rho, Cdc42, Arfs, RABs, RHEB, Vav, Tiam,Sos, Dbl, PRK, TSC1,2, Ras-GAP, Arf-GAPs, Rho-GAPs, caspases, Caspase 2,Caspase 3, Caspase 6, Caspase 7, Caspase 8, Caspase 9, Bcl-2, Mcl-1,Bcl-XL, Bcl-w, Bcl-B, A1, Bax, Bak, Bok, Bik, Bad, Bid, Bim, Bmf, Hrk,Noxa, Puma, IAPB, XIAP, Smac, survivin, Plk1, Cdk4, Cdk 6, Cdk 2, Cdk1,Cdk 7, Cyclin D, Cyclin E, Cyclin A, nucleoside transporters, Ets, Elk,SMADs, Rel-A (p65-NFKB), CREB, NFAT, ATF-2, AFT, Myc, Fos, Sp1, Egr-1,T-bet, β-catenin, HIFs, FOXOs, E2Fs, SRFs, TCFs, Egr-1, β-catenin,STAT1, STAT 3, STAT 4, STAT 5, STAT 6, Cyclin B, Rb, p16, p14Arf,p27KIP, p21CIP, molecular chaperones, Hsp90s, Hsp70, Hsp27, metabolicenzymes, Acetyl-CoA Carboxylase, ATP citrate lyase, nitric oxidesynthase, caveolins, endosomal sorting complex required for transport(ESCRT) proteins, vesicular protein sorting (Vsps), hydroxylases,prolyl-hydroxylases PHD-1, 2 and 3, asparagine hydroxylase FIHtransferases, Pin1 prolyl isomerase, topoisomerases, deacetylases,Histone deacetylases, sirtuins, histone acetylases, CBP/P300 family,MYST family, ATF2, DNA methyl transferases, DMNT1, DMNT3a, DMNT3b,Histone H3K4 demethylases, H3K27, JHDM2A, UTX, VHL, WT-1, p53, Hdm,PTEN, ubiquitin proteases, urokinase-type plasminogen activator (uPA)and uPA receptor (uPAR) system, cathepsins, metalloproteinases,esterases, hydrolases, separase, potassium channels, sodium channels,multi-drug resistance proteins, P-Gycoprotein, p53, WT-1, HMGA, pS6,4EPB-1, eIF4E-binding protein, RNA polymerase, initiation factors,elongation factors.

In some embodiments target analytes may be selected from endogenousanalytes produced by a host or exogenous analytes that are foreign tothe host. Suitable endogenous analytes include, but are not restrictedto, self-antigens that are targets of autoimmune responses as well ascancer or tumour antigens. Illustrative examples of self antigens usefulin the treatment or prevention of autoimmune disorders include, but arenot limited to, antigens associated with diabetes mellitus, arthritis(including rheumatoid arthritis, juvenile rheumatoid arthritis,osteoarthritis, psoriatic arthritis), Crohn's disease, ulcerativecolitis, conjunctivitis, keratoconjunctivitis, ulcerative colitis,asthma, allergic asthma, cutaneous lupus erythematosus, scleroderma,vaginitisμ, proctitis, drug eruptions, leprosy reversal reactions,erythema nodosum leprosum, autoimmune uveitis, allergicencephalomyelitis, multiple sclerosis, myasthenia gravis, systemic lupuserythematosis, autoimmune thyroiditis, dermatitis (including atopicdermatitis and eczematous dermatitis), Wegener's granulomatosis, chronicactive hepatitis, Stevens-Johnson syndrome, idiopathic sprue, lichenplanus, Graves opthalmopathy, sarcoidosis, primary biliary cirrhosis,uveitis posterior, psoriasis, Sjögren's Syndrome, includingkeratoconjunctivitis sicca secondary to Sjögren's Syndrome, alopeciagreata, allergic responses due to arthropod bite reactions, acutenecrotizing haemorrhagic encephalopathy, idiopathic bilateralprogressive sensorineural hearing loss, aplastic anaemia, pure red cellanaemia, idiopathic thrombocytopenia, polychondritis, and interstitiallung fibrosis. Other autoantigens include those derived from nucleosomesfor the treatment of systemic lupus erythematosus. Further non-limitingexamples of analytes include U1-RNP, fibrillin (scleroderma), pancreaticβ cell antigens, GAD65 (diabetes related), insulin, myelin basicprotein, myelin proteolipid protein, histones, PLP, collagen,glucose-6-phosphate isomerase, citrullinated proteins and peptides,thyroid antigens, thyroglobulin, thyroid-stimulating hormone (TSH)receptor, various tRNA synthetases, components of the acetyl cholinereceptor (AchR), MOG, proteinase-3, myeloperoxidase, epidermal cadherin,acetyl choline receptor, platelet antigens, nucleic acids, nucleicacid:protein complexes, joint antigens, antigens of the nervous system,salivary gland proteins, skin antigens, kidney antigens, heart antigens,lung antigens, eye antigens, erythrocyte antigens, liver antigens andstomach antigens.

In some embodiments, the analyte is associated with the presence ofcancer or other tumorous growth. Examples of cancer- and tumor-relatedanalytes detected by binding with an analyte receptor include, but arenot limited to gp100, MART, Melan-A/MART-1, TRP-1, Tyros, TRP2, MC1R,MUC1F, MUC1R, BAGE, GAGE-1, gp100In4, MAGE-1, MAGE-3, MAGE4, PRAME,TRP2IN2, NYNSO1a, NYNSO1b, LAGE1, p97 melanoma antigen, p5 protein,gp75, oncofetal antigen, GM2 and GD2 gangliosides, cdc27, p21ras,gp100^(Pmel117), etv6, aml1, cyclophilin b (acute lymphoblasticleukemia); Imp-1, EBNA-1 (nasopharyngeal cancer); MUC family, HER2/neu,c-erbB-2, MAGE-A4, NY-ESO-1 (ovarian cancer); Prostate Specific Antigen(PSA) and its antigenic epitopes PSA-1, PSA-2, and PSA-3, PSMA,HER2/neu, c-erbB-2, ga733 glycoprotein (prostate cancer); Ig-idiotype (Bcell lymphoma); E-cadherin, α-catenin, β-catenin, γ-catenin, p1120ctn(glioma); p21ras (bladder cancer); p21ras (biliary cancer); HER2/neu,c-erbB-2 (non-small cell lung carcinoma); HER2/neu, c-erbB-2 (renalcancer); viral products such as human papilloma virus proteins (squamouscell cancers of the cervix and oesophagus); NY-ESO-1 (testicularcancer); MUC family, HER2/neu, c-erbB-2 (breast cancer); p53, p21ras(cervical carcinoma); p21ras, HER2/neu, c-erbB-2, MUC family,Cripto-1protein, Pim-1 protein (colon carcinoma); Colorectal associatedantigen (CRC)—CO17-1A/GA733, APC (colorectal cancer); carcinoembryonicantigen (CEA) (colorectal cancer; choriocarcinoma); cyclophilin b(epithelial cell cancer); HER2/neu, c-erbB-2, ga733 glycoprotein(gastric cancer); α-fetoprotein (hepatocellular cancer); Imp-1, EBNA-1(Hodgkin's lymphoma); CEA, MAGE-3, NY-ESO-1 (lung cancer); cyclophilin b(lymphoid cell-derived leukemia); MUC family, p21ras (myeloma); andHTLV-1 epitopes (C cell leukemia).

In some embodiments, the analyte is a foreign antigen. Foreign antigensinclude, but are not limited to, transplantation antigens, allergens,and antigens from pathogenic organisms. Transplantation antigens can bederived from donor cells or tissues from e.g., heart, lung, liver,pancreas, kidney, neural graft components, or from the donorantigen-presenting cells bearing MHC loaded with self antigen in theabsence of exogenous antigen. Non-limiting examples of allergens includeFel d 1 (i.e., the feline skin and salivary gland allergen of thedomestic cat); Der p L Der p II, or Der fi (i.e., the major proteinallergens from the house dust mite); and allergens derived from: grass,tree and weed (including ragweed) pollens; fungi and moulds; foods suchas fish, shellfish, crab, lobster, peanuts, nuts, wheat gluten, eggs andmilk; stinging insects such as bee, wasp, and hornet and the chimomidae(non-biting midges); other insects such as the housefly, fruitfly, sheepblow fly, screw worm fly, grain weevil, silkworm, honeybee, non-bitingmidge larvae, bee moth larvae, mealworm, cockroach and larvae ofTenibrio molitor beetle; spiders and mites, including the house dustmite; allergens found in the dander, urine, saliva, blood or otherbodily fluid of mammals such as cat, dog, cow, pig, sheep, horse,rabbit, rat, guinea pig, mouse and gerbil; airborne particulates ingeneral; latex; and protein detergent additives.

In some embodiments, the analyte is a pathogen or a product or fragmentthereof. Exemplary pathogens include, but are not limited to, viruses,bacteria, prions, protozoans, single-celled organisms, algae, eggs ofpathogenic organisms, microbes, cysts, molds, fungus, worms, amoeba,pathogenic proteins, parasites, algae, and viroids. Many pathogens, andmarkers thereof, are known in the art (see e.g., Foodborne Pathogens:Microbiology and Molecular Biology, Caister Academic Press, eds.Fratamico, Bhunia, and Smith (2005); Maizels et al., Parasite AntigensParasite Genes: A Laboratory Manual for Molecular Parasitology,Cambridge University Press (1991); National Library of Medicine;US20090215157; and US20070207161). Illustrative examples of virusesinclude viruses responsible for diseases including, but not limited to,measles, mumps, rubella, poliomyelitis, hepatitis (e.g. hepatitis A, B,C, delta, and E viruses), influenza, adenovirus, rabies, yellow fever,Epstein-Barr virus and other herpesviruses such as papillomavirus, Ebolavirus, influenza virus, Japanese encephalitis, dengue virus, hantavirus,Sendai virus, respiratory syncytial virus, othromyxoviruses, vesicularstomatitis virus, visna virus, cytomegalovirus, and humanimmunodeficiency virus (HIV). Any suitable antigen derived from suchviruses are useful in the practice of the present invention. Forexample, illustrative retroviral antigens derived from HIV include, butare not limited to, antigens such as gene products of the gag, pol, andenv genes, the Nef protein, reverse transcriptase, and other HIVcomponents. Illustrative examples of herpes simplex viral antigensinclude, but are not limited to, antigens such as immediate earlyproteins, glycoprotein D, and other herpes simplex viral antigencomponents. Non-limiting examples of varicella zoster viral antigensinclude antigens such as 9PI, gpII, and other varicella zoster viralantigen components. Non-limiting examples of Japanese encephalitis viralantigens include antigens such as proteins E, M-E, M-E-NS 1, NS 1, NS1-NS2A, and other Japanese encephalitis viral antigen components.Illustrative examples of hepatitis viral antigens include, but are notlimited to, antigens such as the S, M, and L proteins of hepatitis Bvirus, the pre-S antigen of hepatitis B virus, and other hepatitis(e.g., hepatitis A, B, and C), viral components such as viral DNA and/orRNA. Illustrative examples of influenza viral antigens include; but arenot limited to, antigens such as hemagglutinin and neurarnimidase andother influenza viral components. Illustrative examples of measles viralantigens include, but are not limited to, antigens such as the measlesvirus fusion protein and other measles virus components. Illustrativeexamples of rubella viral antigens include, but are not limited to,antigens such as proteins E1 and E2 and other rubella virus components;rotaviral antigens such as VP7sc and other rotaviral components.Illustrative examples of cytomegaloviral antigens include, but are notlimited to, antigens such as envelope glycoprotein B and othercytomegaloviral antigen components. Non-limiting examples of respiratorysyncytial viral antigens include antigens such as the RSV fusionprotein, the M2 protein and other respiratory syncytial viral antigencomponents. Representative examples of rabies viral antigens include,but are not limited to, antigens such as rabies glycoprotein, rabiesnucleoprotein and other rabies viral antigen components. Illustrativeexamples of papillomavirus antigens include, but are not limited to, theL1 and L2 capsid proteins as well as the E6/E7 antigens associated withcervical cancers. See e.g. Fundamental Virology, Second Edition, eds.Fields, B. N. and Knipe, D. M., 1991, Raven Press, New York, foradditional examples of viral antigens.

Illustrative examples of fungi include Acremoniuin spp., Aspergillusspp., Epidermophytoni spp., Exophiala jeanselmei, Exserohilunm spp.,Fonsecaea compacta, Fonsecaea pedrosoi, Fusarium oxysporum, Basidiobolusspp., Bipolaris spp., Blastomyces derinatidis, Candida spp.,Cladophialophora carrionii, Coccoidiodes immitis, Conidiobolus spp.,Cryptococcus spp., Curvularia spp., Fusarium solani, Geotrichumcandidum, Histoplasma capsulatum var. capsulatum, Histoplasma capsulatumvar. duboisii, Hortaea werneckii, Lacazia loboi, Lasiodiplodiatheobromae, Leptosphaeria senegalenisis, Piedra iahortae, Pityriasisversicolor, Pseudallesheria boydii, Pyrenochaeta romeroi, Rhizopusarrhizus, Scopulariopsis brevicaulis, Scytalidium dimidiatum, Sporothrixschenckii, Trichophyton spp., Trichosporon spp., Zygomcete fungi,Madurella grisea, Madurella mycetomatis, Malassezia furfur, Microsporumspp., Neotestudina rosatii, Onychocola canadensis, Paracoccidioidesbrasiliensis, Phialophora verrucosa, Piedraia hortae, Absidiacoryinbifera, Rhizomucor pusillus, and Rhizopus arrhizus. Thus,illustrative fungal antigens that can be used in the compositions andmethods of the present invention include, but are not limited to,candida fungal antigen components; cryptococcal fungal antigens such ascapsular polysaccharides and other cryptococcal fungal antigencomponents; histoplasma fungal antigens such as heat shock protein 60(HSP60) and other histoplasma fungal antigen components; coccidiodesfungal antigens such as spherule antigens and other coccidiodes fungalantigen components; and tinea fungal antigens such as trichophytin andother coccidiodes fungal antigen components.

Illustrative examples of bacteria include bacteria that are responsiblefor diseases including, but not limited to, diphtheria (e.g.,Corynebacterium diphtheria), pertussis (e.g., Bordetella pertussis),anthrax (e.g., Bacillus anthracia), typhoid, plague, shigellosis (e.g.,Shigella dysenteriae), botulism (e.g., Clostridium botulinum), tetanus(e.g., Clostridium tetani), tuberculosis (e.g., Mycobacteriumtuberculosis), bacterial pneumonias (e.g., Haemophilus influenzae.),cholera (e.g., Vibrio cholerae), salmonellosis (e.g., Salmonella typhi),peptic ulcers (e.g., Helicobacter pylori), Legionnaire's Disease (e.g.Legionella spp.), and Lyme disease (e.g. Borrelia burgdorferi). Otherpathogenic bacteria include Escherichia coli, Clostridium perfringens,Clostridium difficile, Pseudomonas aeruginosa, Staphylococcus aureus,and Streptococcus pyogenes. Further examples of bacteria includeStaphylococcus epidermidis, Staphylococcus sp., Streptococcuspneumoniae, Streptococcus agalactiae, Enterococcus sp., Bacillus cereus,Bifidobacterium bifidum, Lactobacillus sp., Listeria monocytogenes,Nocardia sp., Rhodococcus equi, Erysipelothrix rhusiopathiae,Propionibacterium acnes, Actinomyces sp., Mobiluncus sp.,Peptostreptococcus sp., Neisseria gonorrhoeae, Neisseria meningitides,Moraxella catarrhalis, Veillonella sp., Actinobacillusactinomycetemcomitans, Acinetobacter baumannii, Brucella sp.,Campylobacter sp., Capnocytophaga sp., Cardiobacterium hominis,Eikenella corrodens, Francisella tularensis, Haemophilus ducreyi,Helicobacter pylori, Kingella kingae, Legionella pneumophila,Pasteurella multocida, Klebsiella granulomatis, Enterobacteriaceae,Citrobacter sp., Enterobacter sp., Escherichia coli, Klebsiellapneumoniae, Proteus sp., Salmonella enteriditis, Salmonella typhi,Shigella sp., Serratia marcescens, Yersinia enterocolitica, Yersiniapestis, Aeromonas sp., Plesiomonas shigelloides, Vibrio cholerae, Vibrioparahaemolyticus, Vibrio vulnificus, Acinetobacter sp., Flavobacteriumsp., Burkholderia cepacia, Burkholderia pseudomallei, Xanthomonasmaltophilia, Stenotrophomonas maltophila, Bacteroides fragilis,Bacteroides sp., Prevotella sp., Fusobacterium. sp., and Spirillumminus. Thus, bacterial antigens which can be used in the compositionsand methods of the invention include, but are not limited to: pertussisbacterial antigens such as pertussis toxin, filamentous hemagglutinin,pertactin, F M2, FIM3, adenylate cyclase and other pertussis bacterialantigen components; diphtheria bacterial antigens such as diphtheriatoxin or toxoid and other diphtheria bacterial antigen components;tetanus bacterial antigens such as tetanus toxin or toxoid and othertetanus bacterial antigen components, streptococcal bacterial antigenssuch as M proteins and other streptococcal bacterial antigen components;gram-negative bacilli bacterial antigens such as lipopolysaccharides andother gram-negative bacterial antigen components; Mycobacteriumtuberculosis bacterial antigens such as mycolic acid, heat shock protein65 (HSP65), the 30 kDa major secreted protein, antigen 85A and othermycobacterial antigen components; Helicobacter pylori bacterial antigencomponents, pneumococcal bacterial antigens such as pneumolysin,pneumococcal capsular polysaccharides and other pnermiococcal bacterialantigen components; Haemophilus influenza bacterial antigens such ascapsular polysaccharides and other Haemophilus influenza bacterialantigen components; anthrax bacterial antigens such as anthraxprotective antigen and other anthrax bacterial antigen components;rickettsiae bacterial antigens such as rompA and other rickettsiaebacterial antigen component. Also included with the bacterial antigensdescribed herein are any other bacterial, mycobacterial, mycoplasmal,rickettsial, or chlamydial antigens.

Illustrative examples of protozoa and other parasites that areresponsible for diseases include, but not limited to, malaria (e.g.Plasmodium falciparum), hookworm, tapeworms, helminths, whipworms,ringworms, roundworms, pinworms, ascarids, filarids, onchocerciasis(e.g., Onchocerca volvulus), schistosomiasis (e.g. Schistosoma spp.),toxoplasmosis (e.g. Toxoplasma spp.), trypanosomiasis (e.g. Trypanosomaspp.), leishmaniasis (Leishmania spp.), giardiasis (e.g. Giardialamblia), amoebiasis (e.g. Entamoeba histolytica), filariasis (e.g.Brugia malayi), and trichinosis (e.g. Trichinella spiralis). Thus,antigens which can be used in the compositions and methods of theinvention include, but are not limited to: plasmodium falciparumantigens such as merozoite surface antigens, sporozoite surfaceantigens, circumsporozoite antigens, gametocyte/gamete surface antigens,blood-stage antigen pf 155/RESA and other plasmodial antigen components;leishmania major and other leishmaniae antigens such as gp63,lipophosphoglycan and its associated protein and other leishmanialantigen components; toxoplasma antigens such as SAG-1, p30 and othertoxoplasmal antigen components; schistosomae antigens such asglutathione-S-transferase, paramyosin, and other schistosomal antigencomponents; and trypanosoma cruzi antigens such as the 75-77 kDaantigen, the 56 kDa antigen and other trypanosomal antigen components.

In some embodiments, the analyte is a drug or drug metabolite. A featureof the system is the ability to run any type of assay on the samesystem.

In some embodiments, certain analytes provided herein may be detected ina nucleic acid assay (e.g. a nucleic acid amplification assay). Thesenucleic acid assays may contain one or more nucleic acid probes whichspecifically hybridize with a nucleic acid that is part of or is relatedto an analyte of interest. For example, nucleic acid probes mayspecifically hybridize with a nucleic acid encoding a protein analytedescribed herein. In another example, nucleic acid probes mayspecifically hybridize with a nucleic acid from a pathogen describedherein. These or other nucleic acid probes may be provided, for example,in an assay unit, reagent unit, vessel, tip, or container in a cartridgeor assay station provided herein. Nucleic acid probes may be provided invarious forms, including, for example, in lyophilized, gel, or liquidforms.

Detection

In some embodiments, binding of one or more analyte receptors to one ormore target analytes is detected using one or more detectable labels ortags. In general a label is a molecule that can be directly (i.e., aprimary label) or indirectly (i.e., a secondary label) detected; forexample a label can be visualized and/or measured or otherwiseidentified so that its presence or absence can be known. A label can bedirectly or indirectly conjugated to one or more of an analyte receptor,an analyte, or a tag (e.g. a probe) that interacts with either or bothof the analyte or analyte receptor. In general, a label provides adetectable signal. Non-limiting examples of labels useful in theinvention include fluorescent dyes (e.g., fluorescein isothiocyanate,Texas red, rhodamine, and the like), enzymes (e.g., LacZ, CAT,horseradish peroxidase, alkaline phosphatase, I 2-galactosidase,β-galactosidase, and glucose oxidase, acetylcholinesterase and others,commonly used as detectable enzymes), quantum dot-labels,chromophore-labels, enzyme-labels, affinity ligand-labels,electromagnetic spin labels, heavy atom labels, probes labeled withnanoparticle light scattering labels or other nanoparticles, fluoresceinisothiocyanate (FITC), TRITC, rhodamine, tetramethylrhodamine,R-phycoerythrin, Cy-3, Cy-5, Cy-7, Texas Red, Phar-Red, allophycocyanin(APC), epitope tags such as the FLAG or HA epitope, and enzyme tags suchas and hapten conjugates such as digoxigenin or dinitrophenyl, ormembers of a binding pair that are capable of forming complexes such asstreptavidin/biotin, avidin/biotin or an antigen/antibody complexincluding, for example, rabbit IgG and anti-rabbit IgG; magneticparticles; electrical labels; thermal labels; luminescent molecules;phosphorescent molecules; chemiluminescent molecules; fluorophores suchas umbelliferone, fluorescein, rhodamine, tetramethyl rhodamine, eosin,green fluorescent protein, erythrosin, coumarin, methyl coumarin,pyrene, malachite green, stilbene, lucifer yellow, Cascade Blue,dichlorotriazinylamine fluorescein, dansyl chloride, phycoerythrin,fluorescent lanthanide complexes such as those including Europium andTerbium, molecular beacons and fluorescent derivatives thereof, aluminescent material such as luminol; light scattering or plasmonresonant materials such as gold or silver particles or quantum dots;radiolabels or heavy isotopes including ¹⁴C, ¹²³I, ¹²⁴I, ¹³¹I, ¹²⁵I,Tc99m, ³²P, ³⁵S or ³H; or spherical shells; and probes labeled with anyother signal generating label known to those of skill in the art, asdescribed, for example, in Principles of Fluorescence Spectroscopy,Joseph R. Lakowicz (Editor), Plenum Pub Corp, 2nd edition (July 1999)and the 6 th Edition of the Molecular Probes Handbook by Richard P.Hoagland. Two or more different labels may be used together to detecttwo or more analytes in a single assay. In some embodiments, about ormore than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, 18, 19, 20, or more different labels are used in a single assay.

In some embodiments, the label is an enzyme, the activity of whichgenerates a product having a detectable signal. Substrates used forsensitive detection can be colorimetric, radioactive, fluorescent orchemiluminescent. Conventional colorimetric substrates produce a newcolor (or change in spectral absorption) upon enzyme action on achromogenic substrate. In general, colorimetric substrates produce achange in spectral absorption. In some embodiments, the enzyme ishorseradish peroxidase, substrates of which include but are not limitedto 3,3′-diaminobenzidine (DAB), 3-Amino-9-ethylcarbazole (AEC), andBajoran Purple. In some embodiments, the enzyme is alkaline phosphatase,substrates of which include but are not limited to Fast Red and FerangiBlue. A variety of other enzymatic labels and associated chromagens areknown in the art, and are available from commercial suppliers such asThermo Fisher Scientific. A non-limiting example of an enzymatic assayis an enzyme-linked immunosorbant assay (ELISA). Methods for performingELISA are known in the art, and may be similarly applied in the methodsof the invention. An analayte may or may not be bound by a first analytereceptor that is not labeled before exposure to a second analytereceptor that is labeled (e.g. sandwich ELISA) and specifically binds toeither the analyte or the first analyte receptor. In a typical ELISAassay, the analyte receptor linked to an enzyme is an antibody. Similarassays may be performed where the antibody is replace with anotheranalyte receptor.

Suitable fluorescent labels include, but are not limited to,fluorescein, rhodamine, tetramethylrhodamine, eosin, erythrosin,coumarin, methyl-coumarins, pyrene, Malacite green, stilbene, LuciferYellow, Cascade Blue™, Texas Red, IAEDANS, EDANS, BODIPY FL, LC Red 640,Cy 5, Cy 5.5, LC Red 705 and Oregon green. Suitable optical dyes aredescribed in the 1996 Molecular Probes Handbook by Richard P. Haugland,hereby expressly incorporated by reference. Suitable fluorescent labelsalso include, but are not limited to, green fluorescent protein (GFP),enhanced GFP (EGFP), blue fluorescent protein (BFP), enhanced yellowfluorescent protein (EYFP), luciferase, β-galactosidase, and Renilla.Further examples of fluorescent labels are described in WO 92/15673; WO95/07463; WO 98/14605; WO 98/26277; WO 99/49019; U.S. Pat. No.5,292,658; U.S. Pat. No. 5,418,155; U.S. Pat. No. 5,683,888; U.S. Pat.No. 5,741,668; U.S. Pat. No. 5,777,079; U.S. Pat. No. 5,804,387; U.S.Pat. No. 5,874,304; U.S. Pat. No. 5,876,995; and U.S. Pat. No.5,925,558, which are incorporated herein by reference.

In some embodiments, labels for use in the present invention include:Alexa-Fluor dyes (Alexa Fluor 350, Alexa Fluor 430, Alexa Fluor 488,Alexa Fluor 546, Alexa Fluor 568, Alexa Fluor 594, Alexa Fluor 633,Alexa Fluor 660, Alexa Fluor 680), Cascade Blue, Cascade Yellow andR-phycoerythrin (PE) (Molecular Probes) (Eugene, Oreg.), FITC,Rhodamine, and Texas Red (Pierce, Rockford, Ill.), Cy5, Cy5.5, Cy7(Amersham Life Science, Pittsburgh, Pa.). Tandem conjugate protocols forCy5PE, Cy5.5PE, Cy7PE, Cy5.5APC, Cy7APC are known in the art.Quantitation of fluorescent probe conjugation may be assessed todetermine degree of labeling and protocols including dye spectralproperties are also well known in the art. In some embodiments thefluorescent label is conjugated to an aminodextran linker which isconjugated to a binding element or antibody. Additional labels arelisted in and are available through the on-line and hard copy cataloguesof BD Biosciences, Beckman Coulter, AnaSpec, Invitrogen, Cell SignalingTechnology, Millipore, eBioscience, Caltag, Santa Cruz Biotech, Abcamand Sigma, the contents of which are incorporated herein by reference.

Labels may be associated with the analyte receptor, the analyte, orboth, which association may be covalent or non-covalent. Detection mayresult from either an increase or decrease in a detectable signal from alabel. In some embodiments, the degree of increase or decreasecorrelates with the amount of an analyte. In some embodiments, a samplecontaining analytes to be analyzed is treated with a labeling compoundto conjugate the analytes with a label, such as a fluorescent tag.Binding can then be measured by detection of the label, such as bymeasuring fluorescence, to detect presence and optionally quantity ofone or more analytes, such as in combination with analyte receptorscoupled to an array or analyte receptors coupled to coded beads. In someembodiments, the sample is treated with a labeling compound to conjugatethe analytes with a linker. Upon binding the linker is functionalizedwith a label, such as a fluorescent tag, and the positive event ismeasured by detection of the tag, such as an increase in fluorescence.In some embodiments, the analyte binding domain of an analyte receptoris bound by a probe comprising a label, such as a fluorescent label;upon binding to the analyte, the probe is released, which results in ameasurable decrease in a detectable signal from the label (e.g. adecrease in fluorescence). In some embodiments, an analyte receptor isfluorescently labeled and is partially bound by a probe labeled with aquencher that is in proximity to the fluorescent label; upon binding tothe analyte, the complementary probe is released resulting in ameasurable increase in fluorescence of the label conjugated to theanalyte receptor. In some embodiments, the analyte receptor is bound bya probe, which hybridization occludes a domain containing a secondarystructure; upon binding to the analyte, the probe is released, and thesecondary structure is made available to a label, such as anintercalating dye, used to produce a measurable signal. Labels useful inthe detection of binding between an analyte receptor and an analyte in abinding pair can include, for example, fluorescein,tetramethylrhodamine, Texas Red, or any other fluorescent moleculesknown in the art. The level of label detected will then vary with theamount of target analyte in the mixture being assayed.

In some embodiments, a displaced probe is conjugated to one member of anaffinity pair, such as biotin. A detectable molecule is then conjugatedto the other member of the affinity pair, for example avidin. After atest mixture is applied to an assay unit comprising analyte receptors, adetectable molecule is added. The amount of detectable molecule willvary inversely with the amount of target molecule present in the testmixture. In another embodiment, the displaced probe will be biotinlabeled, and can be detected by addition of fluorescently labeledavidin; the avidin itself will then be linked to another fluorescentlylabeled, biotin-conjugated compound. The biotin group on the displacedoligonucleotide can also be used to bind an avidin-linked reporterenzyme; the enzyme will then catalyze a reaction leading to thedeposition of a detectable compound. Alternatively, the reporter enzymewill catalyze the production of an insoluble product that will locallyquench the fluorescence of an intrinsically-fluorescent solid surface.In another embodiment of a displacement assay, a displaced probe will belabeled with an immunologically-detectable probe, such as digoxigenin.The displaced probe will then be bound by a first set of antibodies thatspecifically recognize the probe. These first antibodies will then berecognized and bound by a second set of antibodies that arefluorescently labeled or conjugated to a reporter enzyme.

In some embodiments, an analyte receptor, such as an antibody, inducesan agglutination reaction in the presence of one or more target analytes(e.g. antigens). Typical agglutination reactions involving the use ofantibodies include (i) mixing polyclonal antibodies with a samplecontaining an antigen corresponding to the antibodies, and observing theformation of immunoagglutinates; (ii) mixing a monoclonal antibody witha sample containing an antigen carrying at least two antigenic functions(bivalent or multivalent antigen) and observing the formation ofimmunoagglutinates; (iii) mixing at least two different monoclonalantibodies with a sample containing a monovalent antigen and observingimmunoagglutination; (iv) any of the reactions mentioned above, butapplying the antibodies, or other suitable analyte receptor as describedherein, coupled to particles, such as latex particles, colloids, etc.;and (v) any of the reactions mentioned above, but applied to antigenspresent on cell surfaces in which case the number of antigens perphysical unit is normally a hundred or more, and in which case suchcells may be agglutinated by monoclonal antibodies, or other suitableanalyte receptor as described herein, even if each antigen molecule ismonovalent. Agglutination reactions can be observed on the surface of asolid substrate such as a glass or plastic plate, or in a solution, suchas in a microtitre plate, cuvette, tip, capillary, or other suitablecontainer. The solid surface or container is preferably colored tocontrast with the color of the agglutinate. In some embodiments, thesolid surface or container is optically clear, such that agglutinationmay be measured by changes in color, contrast, absorbance, or detectionof any other suitable label as described herein. In some embodiments,agglutination is measured is a fluid flow, where the presence of anagglutinate is determined by disruptions in the flow of the fluid. Insome embodiments the agglutination reaction is a hemagglutinationreaction. In some embodiments, the agglutination reaction is anagglutination inhibition reaction, wherein the presence of an analyte(e.g. a small molecule, drug, or drug metabolite) inhibits or slows therate of an agglutination reaction, such as by competing for binding withan analyte receptor (e.g. an antibody) in the presence of anagglutinatable target (e.g. beads coated with analyte).

Receptor binding assays as described herein may be combined with one ormore other assays, such as on different samples within a system of theinvention, or on the same sample. Different assays may be performedsimultaneously or sequentially on one or more samples.

In some embodiments, multiple analytes can assayed simultaneously.Multiple analytes may be analyzed in separate vessels or in the samevessel. The same analyte might be assayed using different detectors.This may allow the system to maintain high precision on differentconcentration ranges of the analyte.

Nucleic Acid Hybridization Assays

In some embodiments, the analyte is a target nucleic acid (e.g. DNA,RNA, mRNA, miRNA, rRNA, tRNA, and hybrids of these) that is detected ina nucleic acid hybridization reaction. Target nucleic acid in a samplemay be a nucleic acid from the subject from which the sample is derived,or from a source to which the subject providing the sample is a host,such as a pathogen as described herein. In general, hybridization refersto a reaction in which one or more polynucleotides react to form acomplex that is stabilized via hydrogen bonding between the bases of thenucleotide residues. The hydrogen bonding may occur by Watson Crick basepairing, Hoogstein binding, or in any other sequence specific manner.The complex may comprise two strands forming a duplex structure, threeor more strands forming a multi stranded complex, a single selfhybridizing strand, or any combination of these. A hybridizationreaction may constitute a step in a more extensive process, such as theinitiation of an amplification process (e.g. PCR, ligase chain reaction,self-sustained sequence replication), or the enzymatic cleavage of apolynucleotide by an endonuclease. A sequence capable of hybridizingwith a given sequence is referred to as the “complement” of the givensequence. In some embodiments, hybridization occurs between a targetnucleic acid (analyte) and a nucleic acid probe. In some embodiments,the target nucleic acid is modified before hybridization with a probe,such as by the ligation of an adapter to one or both ends of the targetnucleic acid to generate a modified target nucleic acid. In a modifiednucleic acid comprising a linker, a probe may hybridize only to linkersequence, only to target nucleic acid sequence, or to both linker andtarget nucleic acid sequence. Non-limiting examples of uses for nucleicacid probes of the invention include detecting the presence of viral orbacterial nucleic acid sequences indicative of an infection, detectingthe presence of variants or alleles of mammalian genes associated withdisease and cancers, genotyping one or more genetic loci (e.g. singlenucleotide polymorphisms), identifying the source of nucleic acids foundin forensic samples, and determining paternity.

The nucleic acid probe of this invention may comprise DNA, RNA, modifiednucleotides (e.g. methylated or labeled nucleotides), modified backbonechemistries (e.g. morpholine ring-containing backbones), nucleotideanalogs, or combinations of two or more of these. The probe can be thecoding or complementary strand of a complete gene or gene fragment, oran expression product thereof. The nucleotide sequence of the probe canbe any sequence having sufficient complementarity to a nucleic acidsequence in a biological sample to allow for hybridization of the probeto the target nucleic acid in the biological sample under a desiredhybridization condition. Ideally, the probe will hybridize only to thenucleic acid target of interest in the sample and will not bindnon-specifically to other non-complementary nucleic acids in the sampleor other regions of the target nucleic acid in the sample. Thehybridization conditions can be varied according to the degree ofstringency desired in the hybridization reaction. For example, if thehybridization conditions are for high stringency, the probe will bindonly to the nucleic acid sequences in the sample with which it has avery high degree of complementarity. Low stringency hybridizationconditions will allow for hybridization of the probe to nucleic acidsequences in the sample which have some complementarity but which arenot as highly complementary to the probe sequence as would be requiredfor hybridization to occur at high stringency. The hybridizationconditions will vary depending on the biological sample, probe type andtarget. An artisan will know how to optimize hybridization conditionsfor a particular application of the present method, or alternatively,how to design nucleic acid probes for optimal use under a specified setof conditions. Also, references herein to “the nucleic acid probe ofthis invention”, “the nucleic acid probe”, and the like may refer to anyof the various embodiments of nucleic acid probes described herein.

The nucleic acid probe can be commercially obtained or can besynthesized according to standard nucleotide synthesizing protocols wellknown in the art. Alternatively, the probe can be produced by isolationand purification of a nucleic acid sequence from biological materialsaccording to methods standard in the art of molecular biology (Sambrooket al. 1989. Molecular Cloning: A Laboratory Manual, 2d Ed., Cold SpringHarbor Laboratory Press, Cold Spring Harbor, N.Y.). The nucleic acidprobe can be amplified according to well known procedure foramplification of nucleic acid (e.g., polymerase chain reaction).Furthermore, the probe of this invention can be linked to any of thelabels of this invention by protocols standard in the art.

It is further contemplated that the present invention also includesmethods for nucleotide hybridization wherein the nucleic acid probe isused as a primer for an enzyme catalyzed elongation reaction such as PCRand primer extension labeling reactions (e.g. in situ and in vitro PCRand other primer extension based reactions). Additionally included aremethods for in situ hybridization.

The labels to which a nucleic acid probe of this invention can be linkedto include, but are not limited to, a hapten, biotin, digoxigenin,fluorescein isothiocyanate (FITC), dinitrophenyl, amino methyl coumarinacetic acid, acetylaminofluorene and mercury-sulfhydryl-ligandcomplexes, chromogenic dyes, fluorescent dyes, and any other suitablelabel as described herein, such as described in combination withlabeling of analyte receptors. In some embodiments, hybridization isdetected indirectly by detection of a product of a hybridizationreaction, such as PCR. For example, amplification products may bedetected by a dye or stain capable of detecting amplified nucleic acids(e.g. intercalating or groove-binding dyes), such as ethidium bromide,SYBR green, SYBR blue, DAPI, acriflavine, fluorcoumanin, ellipticine,daunomycin, chloroquine, distamycin D, chromomycin, propidium iodine,Hoeste, SYBR gold, acridines, proflavine, acridine orange, homidium,mithramycin, ruthenium polypyridyls, anthramycin, and other suitableagents known in the art. In some embodiments, multiple probes, eachhaving a different target nucleic acid and a different label, arehybridized to a single sample simultaneously, such as about or more thanabout 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, or more different probes.

In one embodiment, nucleic acid probes are covalently or non-covalentlycoupled to a substrate. Non-limiting examples of substrates to whichnucleic acid probes may be coupled include microarrays, microbeads,pipette tips, sample transfer devices, cuvettes, capillaries or othertubes, reaction chambers, or any other suitable format compatible withthe subject detection system. Biochip microarray production can employvarious semiconductor fabrication techniques, such as solid phasechemistry, combinatorial chemistry, molecular biology, and robotics. Oneprocess typically used is a photolithographic manufacturing process forproducing microarrays with millions of nucleic acid probes on a singlechip. Alternatively, if the nucleic acid probes are pre-synthesized,they can be attached to an array surface using techniques such asmicro-channel pumping, “ink-jet” spotting, template-stamping, orphotocrosslinking. An exemplary photolithographic process begins bycoating a quartz wafer with a light-sensitive chemical compound toprevent coupling between the quartz wafer and the first nucleotide of aDNA probe being created. A lithographic mask is used to either inhibitor permit the transmission of light onto specific locations of the wafersurface. The surface is then contacted with a solution which may containadenine, thymine, cytosine, or guanine, and coupling occurs only inthose regions on the glass that have been deprotected throughillumination. The coupled nucleotide bears a light-sensitive protectinggroup, allowing the cycle can be repeated. In this manner, themicroarray is created as the probes are synthesized via repeated cyclesof deprotection and coupling. The process may be repeated until theprobes reach their full length. Commercially available arrays aretypically manufactured at a density of over 1.3 million unique featuresper array. Depending on the demands of the experiment and the number ofprobes required per array, each wafer, can be cut into tens or hundredsof individual arrays.

Other methods may be used to produce a coated solid surface with nucleicacid probes attached thereto. A coated solid surface may be aLangmuir-Bodgett film, functionalized glass, germanium, silicon, PTFE,polystyrene, gallium arsenide, gold, silver, membrane, nylon, PVP,polymer plastics, or any other material known in the art that is capableof having functional groups such as amino, carboxyl, Diels-Alderreactants, thiol or hydroxyl incorporated on its surface. These groupsmay then be covalently attached to crosslinking agents, so that thesubsequent binding of the nucleic acid probes and target nucleic acidanalyte can occur in solution without hindrance from the biochip.Typical crosslinking groups include ethylene glycol oligomer, diamines,and amino acids. Alternatively, nucleic acid probes may be coupled to anarray using enzymatic procedures, such as described in US20100240544.

In some embodiments, nucleic acid probes are coupled to the surface of amicrobead. Microbeads useful in coupling to nucleic acid probes areknown in the art, and include magnetic and non-magnetic beads.Microbeads can be labeled with 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or moredyes to facilitate coding of the beads and identification of nucleicacid probes joined thereto. Coding of microbeads can be used todistinguish at least 10, 50, 100, 200, 300, 400, 500, 600, 700, 800,900, 1000, 1500, 2000, 5000, or more different microbeads in a singleassay, each microbead corresponding to a different nucleic acid probeswith specificity for a different target nucleic acid analyte.

In some embodiments, nucleic acid probes are coupled to the surface of areaction chamber, such as a tip. For example, the interior surface of atip may be coated with nucleic acid probes specific for a single targetnucleic acid analyte. Alternatively, the interior surface of a tip maybe coated with two or more different nucleic acid probes specific fordifferent target nucleic acid analytes. When two or more differentnucleic acid probes are coupled to the same interior tip surface, eachof the different nucleic acid probes may be coupled at different knownlocations, such as forming distinct ordered rings or bands at differentpositions along the axis of a tip. In this case, multiple differentnucleic acid analytes may be analyzed in the same sample by drawing asample up a tip and allowing nucleic acid analytes contained in thesample to bind with the nucleic acid probes coated at successivepositions along the tip. Binding events can then be visualized asdescribed herein, with the location of each band in a banding patterncorresponding to a specific known nucleic acid analytes.

In some embodiments, the nucleic acid hybridization reaction is asequencing reaction. Sequencing reactions may proceed directly fromsample nucleic acids, or may involve a pre-amplification step, such asreverse transcription and/or PCR. Sequence analysis usingtemplate-dependent synthesis can include a number of differentprocesses. For example, one of the earliest methods for DNA sequencingwas the four-color chain-termination Sanger sequencing methodology inwhich a population of template molecules is used to create a populationof complementary fragments. Primer extension is carried out in thepresence of the four naturally occurring nucleotides, and with asub-population of dye-labeled terminator nucleotides, e.g.,dideoxyribonucleotides, where each type of terminator (ddATP, ddGTP,ddTTP, ddCTP) includes a different detectable label. As a result, anested set of fragments is created where the fragments terminate at eachnucleotide in the template beyond the primer, and are labeled in amanner that permits identification of the terminating nucleotide. Thenested fragment population is then subjected to size-based separation,e.g., using capillary electrophoresis, and the labels associated witheach different sized fragment is identified to identify the terminatingnucleotide. As a result, the sequence of labels moving past a detectorin the separation system provides a direct readout of the sequenceinformation of the synthesized fragments, and by complementarity, theunderlying template (See, e.g., U.S. Pat. No. 5,171,534, incorporatedherein by reference in its entirety for all purposes).

Other examples of template-dependent sequencing methods includesequence-by-synthesis processes, where individual nucleotides areidentified iteratively, as they are added to the growing primerextension product. In one category of sequencing-by-synthesis, a nucleicacid synthesis complex is contacted with one or more nucleotides underconditions that permit the addition of a single base, and little or noextension beyond that base. The reaction is then interrogated orobserved to determine whether a base was incorporated, and provide theidentity of that base. In a second category of sequencing-by-synthesis,addition of nucleotides to the growing nascent strand are observed inreal-time in an uninterrupted reaction process, e.g., without washsteps.

One example of sequencing-by-synthesis is pyrosequencing, which is aprocess that identifies the incorporation of a nucleotide by assayingthe resulting synthesis mixture for the presence of by-products of thesequencing reaction, namely pyrophosphate. In particular, a primer,polymerase template complex is contacted with a single type ofnucleotide. If that nucleotide is incorporated, the polymerizationreaction cleaves the nucleoside triphosphate between the α and βphosphates of the triphosphate chain, releasing pyrophosphate. Thepresence of released pyrophosphate is then identified using achemiluminescent enzyme reporter system that converts the pyrophosphate,with AMP, into ATP, then measures ATP using a luciferase enzyme toproduce measurable light signals. Where light is detected, the base isincorporated, where no light is detected, the base is not incorporated.Following appropriate washing steps, the various bases are cyclicallycontacted with the complex to sequentially identify subsequent bases inthe template nucleic acid. See, e.g., U.S. Pat. No. 6,210,891,incorporated herein by reference in its entirety for all purposes).

In related processes, the primer/template/polymerase complex isimmobilized upon a substrate and the complex is contacted with labelednucleotides. The immobilization of the complex may be through the primersequence, the template sequence and/or the polymerase enzyme, and may becovalent or noncovalent. In general, preferred aspects, particularly inaccordance with the invention provide for immobilization of the complexvia a linkage between the polymerase or the primer and the substratesurface. A variety of types of linkages are useful for this attachment,including, e.g., provision of biotinylated surface components, usinge.g., biotin-PEG-silane linkage chemistries, followed by biotinylationof the molecule to be immobilized, and subsequent linkage through, e.g.,a streptavidin bridge. Other synthetic coupling chemistries, as well asnon-specific protein adsorption can also be employed for immobilization.In alternate configurations, the nucleotides are provided with andwithout removable terminator groups. Upon incorporation, the label iscoupled with the complex and is thus detectable. In the case ofterminator bearing nucleotides, all four different nucleotides, bearingindividually identifiable labels, are contacted with the complex.Incorporation of the labeled nucleotide arrests extension, by virtue ofthe presence of the terminator, and adds the label to the complex. Thelabel and terminator are then removed from the incorporated nucleotide,and following appropriate washing steps, the process is repeated. In thecase of non-terminated nucleotides, a single type of labeled nucleotideis added to the complex to determine whether it will be incorporated, aswith pyrosequencing. Following removal of the label group on thenucleotide and appropriate washing steps, the various differentnucleotides are cycled through the reaction mixture in the same process.See, e.g., U.S. Pat. No. 6,833,246, incorporated herein by reference inits entirety for all purposes).

In yet a further sequence by synthesis process, the incorporation ofdifferently labeled nucleotides is observed in real time as templatedependent synthesis is carried out. In particular, an individualimmobilized primer/template/polymerase complex is observed asfluorescently labeled nucleotides are incorporated, permitting real timeidentification of each added base as it is added. In this process, labelgroups are attached to a portion of the nucleotide that is cleavedduring incorporation. For example, by attaching the label group to aportion of the phosphate chain removed during incorporation, i.e., a β,γ, or other terminal phosphate group on a nucleoside polyphosphate, thelabel is not incorporated into the nascent strand, and instead, naturalDNA is produced. Observation of individual molecules typically involvesthe optical confinement of the complex within a very small illuminationvolume. By optically confining the complex, one creates a monitoredregion in which randomly diffusing nucleotides are present for a veryshort period of time, while incorporated nucleotides are retained withinthe observation volume for longer as they are being incorporated. Thisresults in a characteristic signal associated with the incorporationevent, which is also characterized by a signal profile that ischaracteristic of the base being added. In related aspects, interactinglabel components, such as fluorescent resonant energy transfer (FRET)dye pairs, are provided upon the polymerase or other portion of thecomplex and the incorporating nucleotide, such that the incorporationevent puts the labeling components in interactive proximity, and acharacteristic signal results, that is again, also characteristic of thebase being incorporated (See, e.g., U.S. Pat. Nos. 6,056,661, 6,917,726,7,033,764, 7,052,847, 7,056,676, 7,170,050, 7,361,466, 7,416,844 andPublished U.S. Patent Application No. 2007-0134128, the full disclosuresof which are hereby incorporated herein by reference in their entiretyfor all purposes). A photodetector could be used instead of a CCD camerato detect a change in scattering. A combination of fluorescence andtransmittance can be used to enhance the signal.

Nucleic acid hybridization assays as described herein may be combinedwith one or more other assays, such as on different samples within asystem of the invention, or on the same sample. Different assays may beperformed simultaneously or sequentially on one or more samples.

Various samples may be used for nucleic acid assays provided herein. Forexample, a nasopharyngeal swab, nasopharyngeal aspirate, or sputumsample may be used as a biological sample from which an infectiouspathogen may be detected. In some embodiments, any other biologicalsample described elsewhere herein may be used for a nucleic acid assay.

Nucleic acid assays may be monitored using various method for assaydetection provided herein. For example, nucleic acid amplificationassays may be measured by monitoring the increase in fluorescence of thereaction (for example, in assays in which a fluorescent dye whichintercalculates with double-stranded DNA is used) or by monitoring theincrease in absorbance or turbidity of the reaction (for example, inassays in which the pyrophosphate that is generated, as a result ofnucleotide incorporation during DNA synthesis, reacts with Mg++ to forminsoluble magnesium pyrophosphate). Nucleic acid amplification assaysmay be further analyzed, for example, by obtaining fluorescence,absorbance, or turbidity values over a period of time, and analyzing thedata to identify an inflection point indicating the presence or amountof a nucleic acid of interest in a sample This analysis maybe performed,for example, by fitting the data to an exponential curve and selectingthe inflection point based on a threshold value above baseline. Abaseline may be determined, for example, by using a moving average of atleast 3 data points. An inflection point for data may be selected for aparticular assay, and may be, for example, a time when a value is 10,20, 30, 40, 50, 60, 70, 80, 90, or 100% above a baseline.

General Chemistry Assays

In some embodiments, devices and systems provided herein may beconfigured to perform one or more general chemistry assays. Generalchemistry assays may include, for example, assays of a Basic MetabolicPanel [glucose, calcium, sodium (Na), potassium (K), chloride (Cl), CO2(carbon dioxide, bicarbonate), creatinine, blood urea nitrogen (BUN)],assays of an Electrolyte Panel [sodium (Na), potassium (K), chloride(Cl), CO2 (carbon dioxide, bicarbonate)], assays of a Chem 14Panel/Comprehensive Metabolic Panel [glucose, calcium, albumin, totalprotein, sodium (Na), potassium (K), chloride (Cl), CO2 (carbon dioxide,bicarbonate), creatinine, blood urea nitrogen (BUN), alkalinephosphatase (ALP), alanine aminotransferase (ALT/GPT), aspartateaminotransferase (AST/GOT), total bilirubin] assays of a LipidProfile/Lipid Panel [LDL cholesterol, HDL cholesterol, totalcholesterol, and triglycerides], assays of a Liver Panel/Liver Function[alkaline phosphatase (ALP), alanine aminotransferase (ALT/GPT),aspartate aminotransferase (AST/GOT), total bilirubin, albumin, totalprotein, gamma-glutamyl transferase (GGT), lactate dehydrogenase (LDH),prothrombin time (PT)], alkaline phosphatase (APase), hemoglobin, VLDLcholesterol, ethanol, lipase, pH, zinc protoporphyrin, direct bilirubin,blood typing (ABO, RHD), lead, phosphate, hemagglutination inhibition,magnesium, iron, iron uptake, fecal occult blood, and others,individually or in any combination.

In general chemistry assays provided herein, in some examples, the levelof an analyte in a sample is determined through one or more assay stepsinvolving a reaction of the analyte of interest with one or morereagents, leading to a detectable change in the reaction (e.g. change inthe turbidity of the reaction, generation of luminescence in thereaction, change in the color of the reaction, etc.). In some examples,a property of a sample is determined through one or more assay stepsinvolving a reaction of the sample of interest with one or morereagents, leading to a detectable change in the reaction (e.g. change inthe turbidity of the reaction, generation of luminescence in thereaction, change in the color of the reaction, etc.). Typically, as usedherein, “general chemistry” assays do not involve amplification ofnucleic acids, imaging of cells on a microscopy stage, or thedetermination of the level of an analyte in solution based on the use ofa labeled antibody/binder to determine the level of an analyte in asolution. In some embodiments, general chemistry assays are performedwith all reagents in a single vessel—i.e. to perform the reaction, allnecessary reagents are added to a reaction vessel, and during the courseof the assay, materials are not removed from the reaction or reactionvessel (e.g. there is no washing step; it is a “mix and read” reaction).General chemistry assays may also be, for example, colorimetric assays,enzymatic assays, spectroscopic assays, turbidimetric assays,agglutination assays, coagulation assays, and/or other types of assays.Many general chemistry assays may be analyzed by measuring theabsorbance of light at one or more selected wavelengths by the assayreaction (e.g. with a spectrophotometer). In some embodiments, generalchemistry assays may be analyzed by measuring the turbidity of areaction (e.g. with a spectrophotometer). In some embodiments, generalchemistry assays may be analyzed by measuring the chemiluminescencegenerated in the reaction (e.g. with a PMT, photodiode, or other opticalsensor). In some embodiments, general chemistry assays may be performedby calculations, based on experimental values determined for one or moreother analytes in the same or a related assay. In some embodiments,general chemistry assays may be analyzed by measuring fluorescence of areaction (e.g. with a detection unit containing or connected to i) alight source of a particular wavelength(s) (“excitation wavelength(s)”);and ii) a sensor configured to detect light emitted at a particularwavelength(s) (“emission wavelength(s)”). In some embodiments, generalchemistry assays may be analyzed by measuring agglutination in areaction (e.g. by measuring the turbidity of the reaction with aspectrophotometer or by obtaining an image of the reaction with anoptical sensor). In some embodiments, general chemistry assays may beanalyzed by imaging the reaction at one or more time points (e.g. with aCCD or CMOS optical sensor), followed by image analysis. Optionallyanalysis may involve prothrombin time, activated partial thromboplastintime (APTT), either of which may be measured through a method such asbut not limited to turbidimetry. In some embodiments, general chemistryassays may be analyzed by measuring the viscosity of the reaction (e.g.with a spectrophotometer, where an increase in viscosity of the reactionchanges the optical properties of the reaction). In some embodiments,general chemistry assays may be analyzed by measuring complex formationbetween two non-antibody reagents (e.g. a metal ion to a chromophore;such a reaction may be measured with a spectrophotometer or throughcolorimetry using another device). In some embodiments, generalchemistry assays may be analyzed by non-ELISA or cytometry-based methodsfor assaying cellular antigens (e.g. hemagglutination assay for bloodtype, which may be measured, for example, by turbidity of the reaction).In some embodiments, general chemistry assays may be analyzed with theaid of electrochemical sensors (e.g. for carbon dioxide or oxygen).Additional methods may also be used to analyze general chemistry assays.

In some embodiments, a spectrophotometer may be used to measure ageneral chemistry assay. In some embodiments, general chemistry assaysmay be measured at the end of the assay (an “end-point” assay) or at twoor more times during the course of the assay (a “time-course” or“kinetic” assay).

A glucose assay may be performed, for example, by incubating abiological sample (e.g. plasma, urine, etc.) with glucose oxidase, togenerate gluconic acid and hydrogen peroxide. In an example, thehydrogen peroxide may be incubated with a peroxidase and a chromogenthat can change color when oxidized—e.g. o-dianisidine. A coloredproduct may be further stabilized by reaction with sulfuric acid. Thecolored product may be measured in a spectrophotometer by absorbance at405 nm. In another example, the hydrogen peroxide may be incubated witha peroxidase, 4-aminoantipyrine, and a phenolic compound (e.g. N, Ndiethylaniline), to form a colored product. The product may be measured,for example, in a spectrophotometer at 510 nm.

An alanine aminotransferase assay may be performed, for example, byincubating a biological sample (e.g. plasma, urine, etc.) withalpha-ketoglutarate and alanine, to generate glutamate and pyruvate.Incubation of these products with an oxidizable chromogen [e.g.10-acetyl-3,7-dihydroxyphenoxazine (ADHP)] may generate a coloredproduct. Oxidized 10-acetyl-3,7-dihydroxyphenoxazine may be detected,for example, colorimetrically in a spectrophotometer by absorbance at570 nm, or fluorescently, at EX/EM=535/587 nm. In another example, theglutamate and pyruvate products may be incubated with lactatedehydrogenase and NADH, where pyruvate reacts with NADH in a lactatedehydrogenase-catalyzed reaction to form NAD+ and lactate. This reactionmay be monitored by absorbance at 340 nm, at which NADH absorbs (i.e.the more NADH is consumed, the lower the absorbance at 340 nm).

A potassium assay may be performed, for example, by incubating abiological sample (e.g. plasma, urine, etc.) with tetraphenylborate.Potassium in the plasma may form an insoluble salt with thetetraphenylborate, which may precipitate out of solution and/or increasethe turbidity of the sample. The assay may be measured in aspectrophotometer, for example, by measuring absorbance of the sample at578 nm.

An alkaline phosphatase assay may be performed, for example, byincubating a biological sample (e.g. plasma, urine, etc.) with thechromogen p-nitrophenyl phosphate (pNPP). pNPP may be dephosphorylatedby alkaline phosphatase to form p-nitrophenol and phosphate; it forms ayellow color upon dephosphorylation. The assay may be measured, forexample, in a spectrophotometer by measuring, colorimetrically, theabsorbance of the sample at 405 nm. In another example, an alkalinephosphatase assay may be performed, for example, by incubating plasmawith a chemiluminescent substrate that releases light upon alkalinephosphatase-mediated cleavage (e.g.3-(2′-spiroadamantyl)-4-methoxy-4-(3″-phosphoryloxy)-phenyl-1,2-dioxetane(AMPPD)). The assay may be measured, for example, by obtaining a readingof the assay at a light sensor (e.g. a PMT or photodiode).

A sodium assay may be performed, for example, by incubating a biologicalsample (e.g. plasma, urine, etc.) with beta-galactosidase andortho-nitrophenyl-beta-galactoside (ONPG). Beta-galactosidase may havesodium-dependent activity, and hydrolyze ONPG to galactose andortho-nitrophenol. Ortho-nitrophenol generation may be measured in aspectrophotometer by absorbance at 420 nm.

A calcium assay may be performed, for example, by incubating abiological sample (e.g. plasma, urine, etc.) with o-cresolphthalein and2-amino-2-methyl-1-propanol. Calcium in the plasma may form a complexwith the o-cresolphthalein, the complex having a purple color. Thecomplex may be measured in a spectrophotometer by absorbance at 575 nm.

A hemoglobin assay may be performed, for example, by incubating wholeblood with one or more detergents, ferricyanide, and cyanide. Hemoglobinmay form a complex with cyanide, which may be measured in aspectrophotometer by absorbance at 540 nm.

An HDL-cholesterol assay may be performed, for example, by incubating abiological sample (e.g. plasma, etc.) with reagents that protect non-HDLcholesterol (e.g. LDL, VLDL, and chylomicrons) but leave HDL-cholesterolexposed to enzymes. These reagents may include, for example,polyvinylsulfonic acid (PVS), polyethylene glycol methylether (PEGME) ordextran sulfate. The reaction mixture is then incubated with cholesterolesterase (to convert cholesteryl ester to cholesterol) and cholesteroloxidase (to convert cholesterol to cholest-4-ene-3-one, andsimultaneously producing hydrogen peroxide). The reaction mixture isincubated with an oxidizable chromogen (e.g.N-(2-hydroxy-3-sulfopropyl)-3,5-dimethyoxyaniline (ALPS) andaminoantipyrene (AAP)) or fluorescent dye, which may be oxidized byhydrogen peroxide, catalyzed by a peroxidase (e.g. horseradishperoxidase).

A VLDL-cholesterol assay may be performed, for example, by calculatingthe level of VLDL in a sample based on the enzyme-based determination ofthe level of other cholesterol molecules in the sample (e.g. totalcholesterol and HDL-cholesterol). In some instances, VLDL is estimatedto be one-fifth of the total triglycerides in the sample. In anotherexample, VLDL-cholesterol may be determined with LDL-cholesterol, byphysically or chemically separating LDL and VLDL-cholesterol fromHDL-cholesterol. The isolated LDL/VLDL may then be incubated withcholesterol esterase (to convert cholesteryl ester to cholesterol) andcholesterol oxidase (to convert cholesterol to cholest-4-ene-3-one, andsimultaneously producing hydrogen peroxide). The reaction mixture isincubated with an oxidizable chromogen (e.g. ALPS/AAP) or fluorescentdye, which may be oxidized by hydrogen peroxide, catalyzed by aperoxidase (e.g. horseradish peroxidase).

A pH assay may be performed, for example, by incubating a biologicalsample (e.g. plasma, urine, etc.) with a pH indicator molecule.Commonly, the color of pH indicator changes in response to the pH of thesurrounding solution. pH indicators are well known in the art, andinclude, for example bromophenol blue, methyl red, litmus,phenolphthalein and phenol red.

A prothrombin time (PT) assay may be performed, for example, byincubating blood with citrate or other anticoagulant, and isolating theblood plasma. A substance is then added to the plasma to reverse theeffects of the anticoagulant (e.g. in the case of citrate, calcium isadded). Then tissue factor (factor (III)) is added to the plasma, andthe time required for the sample to clot is measured. Clotting of thesample may, for example, increase the turbidity of the sample and/orincrease its viscosity. The assay may be measured in aspectrophotometer, for example, by measuring absorbance of the sample.

A zinc protoporphyrin assay may be performed, for example, by incubatingred blood cells with a solution to lyse the red blood cells (e.g. wateror water containing 10 mM phosphate buffer, pH 7.4), and observing thefluorescence of the sample at EX/EM=424/594 nm, which are strongexcitation and emission wavelengths for zinc protoporphyrin.

A chloride assay may be performed, for example, by incubating abiological sample (e.g. plasma, urine, etc.) with a reagent which hasone color when in complex with a mercury atom, and which has a secondcolor when in complex with an iron atom (e.g.2,4,6-Tripyridyl-s-triazine (TPTZ) or thiocyanate). These reagentspreferentially form complexes with mercury atoms over iron atoms.However, in the presence of chloride ions, mercury disassociates fromthe reagent and forms HgCl2, and iron is able to form a complex with thereagent. The iron-containing complex may be colored and can be measured,for example, in a spectrophotometer: Fe-TPTZ at 620 nm or Fe-thiocyanateat 480 nm.

A triglycerides assay may be performed, for example, by incubating abiological sample (e.g. plasma, etc.) with lipase enzyme, which canconvert triglycerides to glycerol and fatty acids. Glycerol may then beincubated with additional enzymes which react with glycerol and productsthereof, ultimately resulting in the formation of hydrogen peroxide (inan example, glycerol kinase catalyzes the reaction of glycerol and ATPto form glycerol phosphate, and glycerol-3-phosphate oxidase catalyzesthe conversion of glycerol phosphate to dihydroxyacetone phosphate andhydrogen peroxide). The reaction may be incubated with a peroxidase andoxidizable substrate (e.g. a chromogen or fluorescent dye), theoxidation of which may be monitored (for example, with aspectrophotometer).

A total cholesterol assay may be performed, for example by incubating abiological sample (e.g. plasma, etc.) with cholesterol esterase (toconvert cholesteryl ester to cholesterol) and cholesterol oxidase (toconvert cholesterol to cholest-4-ene-3-one, and simultaneously producinghydrogen peroxide). The reaction may be incubated with a peroxidase andan oxidizable substrate (e.g. a chromogen or fluorescent dye), theoxidation of which may be monitored (for example, with aspectrophotometer).

An albumin assay may be performed, for example, by incubating abiological sample (e.g. plasma, urine, etc.) with a dye which binds toalbumin, such as 5′,5″-dibromo-o-cresolsulfophthalein (BromocresolPurple (BCP)) or Bromocresol Green (BCG)). The assays can be measured,for example, in a spectrophotometer by absorbance at 600 nm for BCP, orabsorbance at 628 nm for BCG.

A total protein assay may be performed, for example, by incubating abiological sample (e.g. plasma, urine, etc.) with a reagent which bindsto one or more structures in proteins (e.g. peptide bonds). Thesereagents include, for example, copper (II) ions (for the Biuret test)and Coomassie™ dyes. Protein-dye complexes may be further stabilized byincubating the complexes with another reagent, such as bicinchoninicacid (BCA) (for the BCA test). For assays with copper (II) ions, thesamples can be measured, for example in a spectrophotometer at 540 nm.For assays with a Coomassie™ dye, the samples can be measured, forexample in a spectrophotometer at 595 nm.

A bicarbonate/carbon dioxide assay may be performed, for example, byadjusting the pH of biological sample (e.g. plasma, urine, etc.) to a pHgreater than 7, so that carbon dioxide in the sample is converted tobicarbonate (HCO3-). Phosphoenolpyruvate (PEP) and phosphoenolpyruvatecarboxylase (PEPC) are provided to the sample, such that PEPC catalyzesthe reaction between PEP and bicarbonate to form oxaloacetate andphosphate. The oxaloacetate may be detected by a variety of mechanisms.For example, oxaloacetate may be incubated with NADH and malatedehydrogenase, in which malate dehydrogenase catalyzes the conversion ofoxaloacetate and NADH to malate and NAD+. The reaction may be monitoredby measuring absorbance at 340 nm, to monitor the level of NADH. Inanother example, oxaloacetate may be incubated with a chromogen whichcan form a complex with oxaloacetate, such as Fast Violet B. Thereaction may be monitored, for example, in a spectrophotomer measuringthe absorbance at 578 nm, to monitor the level of oxaloacetate-FastViolet B complex.

An aspartate aminotransferase (AST/SGOT) assay may be performed, forexample, by incubating a biological sample (e.g. plasma, urine, etc.)with one or more substrates for aspartate aminotransferase (e.g.aspartate and alpha-ketoglutarate). Aspartate aminotransferase in thesample may catalyze the transfer of an amino group from aspartate toalpha-ketoglutarate, to form oxaloacetate and glutamate. Malatedehydrogenase may also be provided in the assay, which may catalyze theconversion of oxaloacetate to malate, coupled with the oxidation of NADHto NAD+. Lactate dehydrogenase may also be provided in the assay, toreduce interference from pyruvate. The assay may be monitored byabsorbance at 340 nm, at which NADH absorbs (i.e. the more NADH isconsumed, the lower the absorbance at 340 nm). The rate of conversion ofNADH to NAD+ may be directly proportional to the quantity of aspartateaminotransferase in the sample.

A blood urea nitrogen (BUN) assay may be performed, for example, byincubating a biological sample (e.g. plasma, urine, etc.) with urease,which may cleave urea in the sample to yield carbon dioxide and ammonia.The ammonia may then be involved in a reaction which may be readilymonitored. In an example, the ammonia may be incubated with ammonia andalpha-ketoglutarate in the presence of glutamate dehydrogenase and NADH,to yield glutamate and NAD+. The assay may be monitored by absorbance at340 nm, at which NADH absorbs; the rate of conversion of NADH to NAD+may be directly proportional to the quantity of urea in the sample. Inanother example, ammonia may be incubated with salicylate and sodiumnitroprusside, and then with hypochlorite, to yield a blue-green coloredproduct, which may be measured, for example, in a spectrophotometer at630 nm.

A total bilirubin assay may be performed, for example, by incubating abiological sample (e.g. plasma, urine, etc.) with a reagent thatconverts bilirubin to a readily detectable molecule. For example, asample may be incubated with sulfanilic acid and sodium nitrite toconvert bilirubin to an azobilirubin form which may, for example, bereadily detected in a spectrophotometer by absorbance at 550 nm.

A creatinine assay may be performed, for example, by incubating abiological sample (e.g. plasma, urine, etc.) with creatininase, whichthat converts creatinine to creatine. Creatine may then be converted tosarcosine by creatine amidinolydrolase. Sarcosine oxidase may thencatalyze the reaction of sarcosine with water and oxygen to formglycine, formaldehyde, and hydrogen peroxide. The hydrogen peroxide maybe used with any oxidizable chromogen or other detectable agentdescribed herein, according to methods disclosed elsewhere herein. Thelevel of the oxidized product may be directly proportional to thequantity of creatinine in the sample.

It should be understood that for substantially all of the foregoing,embodiments of the system herein may process two or more of these assaysin the system using multiple same or different detection methods usingthe same system. This simultaneous processing of at least two,optionally three assays, within the same device (or optionally using thesame system) using aliquots from the same sample provides substantialadvantages over known systems due to savings in at least the reducedamount of sample used and the reduced processing time for the multipleassays.

Electrophoresis

In some embodiments, a system of the invention comprises subjectinganalytes to an electrophoresis process. The present invention may beused for the separation, detection and measurement of one or moreanalytes in one or more samples of biological, ecological, or chemicalinterest. Of particular interest are macromolecules such as proteins,polypeptides, saccharides and polysaccharides, genetic materials such asnucleic acids and polynucleotides, carbohydrates, cellular materialssuch as bacteria, viruses, organelles, cell fragments, metabolites,drugs, any other analyte as described herein, and combinations thereof.Proteins that are of interest include proteins that are present in bloodplasma, albumin, globulin, fibrinogen, blood clotting factors, hormones,interferons, enzymes, growth factors, and other proteins describedherein. Other chemicals that can be separated and detected using thepresent invention include, but are not limited to pharmaceuticals suchas antibiotics, as well as agricultural chemicals such as insecticidesand herbicides.

Electrophoresis may comprise the use of gels and/or capillaries.Electrophoretic separation can be conducted with or without using amolecular matrix (also referred to herein as a sieving matrix or mediumas well as a separation matrix or medium) to effect separation. Where nomatrix is used as part of a capillary electrophoresis process, thetechnique is commonly termed capillary zone electrophoresis (CZE). Wherea matrix is used in combination with a capillary electrophoresisprocess, the technique is commonly termed capillary gel electrophoresis(CGE). Non-limiting examples of matrices for use in electrophersisprocesses include linear polymer solutions, such as apoly(ethyleneoxide) solution, cross-linked polyacrylamide, and agarose.Suitable matrices can be in the form of liquid, gel, or granules.

In electrophoresis, the separation buffer is typically selected so thatit aids in the solubilization or suspension of the species that arepresent in the sample. Typically the liquid is an electrolyte whichcontains both anionic and cationic species. Preferably the electrolytecontains about 0.005-10 moles per liter of ionic species, morepreferably about 0.01-0.5 mole per liter of ionic species. Examples ofan electrolyte for a typical electrophoresis system include mixtures ofwater with organic solvents and salts. Representative materials that canbe mixed with water to produce appropriate electrolytes includesinorganic salts such as phosphates, bicarbonates and borates; organicacids such as acetic acids, propionic acids, citric acids, chloroaceticacids and their corresponding salts and the like; alkyl amines such asmethyl amines; alcohols such as ethanol, methanol, and propanol; polyolssuch as alkane diols; nitrogen containing solvents such as acetonitrile,pyridine, and the like; ketones such as acetone and methyl ethyl ketone;and alkyl amides such as dimethyl formamide, N-methyl and N-ethylformamide, and the like. The above ionic and electrolyte species aregiven for illustrative purposes only. A researcher skilled in the art isable to formulate electrolytes from the above-mentioned species andoptionally species such an amino acids, salts, alkalis, etc., to producesuitable support electrolytes for using capillary electrophoresissystems. The voltage used for electrophoretic separations is notcritical to the invention, and may vary widely. Typical voltages forcapillary electrophoresis are about 500 V-30,000 V, preferably about1,000-20,000 V.

In some embodiments, the electrophoresis process is a capillaryelectrophoresis process. In a typical capillary electrophoresis process,a buffer-filled capillary is suspended between two reservoirs filledwith buffer. An electric field is applied across the two ends of thecapillary. The electrical potential that generates the electric field isin the range of kilovolts. Samples containing one or more components orspecies are typically introduced at the high potential end and under theinfluence of the electrical field. Alternatively, the sample is injectedusing pressure or vacuum. The same sample can be introduced into manycapillaries, or a different sample can be introduced into eachcapillary. Typically, an array of capillaries is held in a guide and theintake ends of the capillaries are dipped into vials that containsamples. After the samples are taken in by the capillaries, the ends ofthe capillaries are removed from the sample vials and submerged in abuffer which can be in a common container or in separate vials. Thesamples migrate toward the low potential end. During the migration,components of the sample are electrophoretically separated. Afterseparation, the components are detected by a detector. Detection may beeffected while the samples are still in the capillaries or after theyhave exited the capillaries.

The channel length for capillary electrophoresis is selected such thatit is effective for achieving proper separation of species. Generally,the longer the channel, the greater the time a sample will take inmigrating through the capillary. Thus, the species may be separated fromone another with greater distances. However, longer channels contributeto the band broadening and lead to excessive separation time. Generally,for capillary electrophoresis, the capillaries are about 10 cm to about5 meters long, and preferably about 20 cm to about 200 cm long. Incapillary gel electrophoresis, where typically a polymer separationmatrix is used, the more preferred channel length is about 10 cm toabout 100 cm long.

The internal diameter (i.e., bore size) of the capillaries is notcritical, although small bore capillaries are more useful in highlymultiplexed applications. The invention extends to a wide range ofcapillary sizes. In general, capillaries can range from about 5-300micrometers in internal diameter, with about 20-100 micrometerspreferred. The length of the capillary can generally range from about100-3000 mm, with about 300-1000 mm preferred.

A suitable capillary is constructed of material that is sturdy anddurable so that it can maintain its physical integrity through repeateduse under normal conditions for capillary electrophoresis. It istypically constructed of nonconductive material so that high voltagescan be applied across the capillary without generating excessive heat.Inorganic materials such as quartz, glass, fused silica, and organicmaterials such as polytetrafluoroethylene, fluorinatedethylene/propylene polymers, polyfluoroethylene, aramide, nylon (i.e.,polyamide), polyvinyl chloride, polyvinyl fluoride, polystyrene,polyethylene and the like can be advantageously used to makecapillaries.

Where excitation and/or detection are effected through the capillarywall, a particularly advantageous capillary is one that is constructedof transparent material, as described in more detail below. Atransparent capillary that exhibits substantially no fluorescence, i.e.,that exhibits fluorescence lower than background level, when exposed tothe light used to irradiate a target species is especially useful incases where excitation is effected through the capillary wall. One sucha capillary is available from Polymicro Technologies (Phoenix, Ariz.).Alternatively, a transparent, non-fluorescing portion can be formed inthe wall of an otherwise nontransparent or fluorescing capillary so asto enable excitation and/or detection to be carried out through thecapillary wall. For example, fused silica capillaries are generallysupplied with a polyimide coating on the outer capillary surface toenhance its resistance to breakage. This coating is known to emit abroad fluorescence when exposed to wavelengths of light under 600 nm. Ifa through-the-wall excitation scheme is used without first removing thiscoating, the fluorescence background can mask a weak analyte signal.Thus, a portion of the fluorescing polymer coating can be removed by anyconvenient method, for example, by boiling in sulfuric acid, byoxidation using a heated probe such as an electrified wire, or byscraping with a knife. In a capillary of approximately 0.1 mm innerdiameter or less, a useful transparent portion is about 0.01 mm to about1.0 mm in width.

Coagulation Assay

In some embodiments a system of the invention comprises subjectinganalytes to a coagulation assay. Coagulation assays include, but are notlimited to, assays for the detection of one or more coagulation factorsand measurement of clotting time. Typically the read-out of acoagulation assay is the formation of a clot, a rate of clot formation,or the time to clot formation. Clotting factors include factor I(fibrinogen), factor II (prothrombin), factor III (tissuethromboplastin), factor IV (calcium), factor V (proaccelerin), factor VI(no longer considered active in hemostasis), factor VII (proconvertin),factor VIII (antihemophilic factor), factor IX (plasma thromboplastincomponent; Christmas factor), factor X (stuart factor), factor XI(plasma thromboplastin antecedent), factor XII (hageman factor), andfactor XIII (fibrin stabilizing factor). Diagnosis of hemorrhagicconditions such as hemophilia, where one or more of the twelve bloodclotting factors may be defective, can be achieved by a wide variety ofcoagulation tests. In addition, several tests have been developed tomonitor the progress of thrombolytic therapy. Other tests have beendeveloped to signal a prethrombolytic or hypercoagulable state, ormonitor the effect of administering protamine to patients duringcardiopulmonary bypass surgery. Coagulation tests are also useful inmonitoring oral and intravenous anticoagulation therapy. Three examplesof diagnostic coagulation tests useful in the present invention areactivated partial thromboplastin time (APTT), prothrombin time (PT), andactivated clotting time (ACT).

An APTT test evaluates the intrinsic and common pathways of coagulation.For this reason APTT is often used to monitor intravenous heparinanticoagulation therapy. Specifically, it measures the time for a fibrinclot to form after the activating agent, such as calcium, and aphospholipid have been added to a citrated blood sample. Heparinadministration has the effect of suppressing clot formation.

A PT test evaluates the extrinsic and common pathways of coagulation(e.g. conversion of prothrombin to thrombin in the presence of calciumions and tissue thromoplastin) and can be used to monitor oralanticoagulation therapy. The oral anticoagulant coumadin suppresses theformation of prothrombin. Consequently, the test is based on theaddition of calcium and tissue thromboplastin to the blood sample.

An ACT test evaluates the intrinsic and common pathways of coagulation.It is often used to monitor anticoagulation via heparin therapy. The ACTtest is based on addition of an activator to the intrinsic pathway tofresh whole blood to which no exogenous anticoagulant has been added.

Coagulation assays may use a turbidimetric method of measurement. In oneexample of coagulation assay analysis, whole-blood samples are collectedinto a citrate vacutainer and then centrifuged. The assay is performedwith plasma to which a sufficient excess of calcium has been added toneutralize the effect of citrate. For a PT test, tissue thromboplastinis provided as a dry reagent that is reconstituted before use. Thisreagent is thermally sensitive and is maintained at 4° C. by theinstruments. Aliquots of sample and reagent are transferred to a cuvetteheated at 37° C., and the measurement is made based on a change inoptical density.

As an alternative to the turbidimetric method, Beker et al. (See,Haemostasis (1982) 12:73) introduced a chromogenic PT reagent(Thromboquant PT). The assay is based on the hydrolysis ofp-nitroaniline from a modified peptide, Tos-Gly-Pro-Arg-pNA, by thrombinand is monitored spectrophotometrically. Coagulation may also bemeasured by changes or disruptions in the flow of a fluid, such as byreduced flow rate, increased flow time between two points, and formationof a blockage to fluid flow, such as in a capillary. Standards fornormal coagulation results to which a test result may be compared willvary with the method used, and are known in the art or may be determinedusing a control sample (e.g. from a normal subject). Cytometry

In some embodiments, the assay system is configured to perform cytometryassays. Cytometry assays are typically used to optically, electrically,or acoustically measure characteristics of individual cells. For thepurposes of this disclosure, “cells” may encompass non-cellular samplesthat are generally of similar sizes to individual cells, including butnot limited to vesicles (such as liposomes), small groups of cells,virions, bacteria, protozoa, crystals, bodies formed by aggregation oflipids and/or proteins, and substances bound to small particles such asbeads or microspheres. Such characteristics include but are not limitedto size; shape; granularity; light scattering pattern (or opticalindicatrix); whether the cell membrane is intact; concentration,morphology and spatio-temporal distribution of internal cell contents,including but not limited to protein content, protein modifications,nucleic acid content, nucleic acid modifications, organelle content,nucleus structure, nucleus content, internal cell structure, contents ofinternal vesicles (including pH), ion concentrations, and presence ofother small molecules such as steroids or drugs; and cell surface (bothcellular membrane and cell wall) markers including proteins, lipids,carbohydrates, and modifications thereof. By using appropriate dyes,stains, or other labeling molecules either in pure form, conjugated withother molecules or immobilized in, or bound to nano- or micro-particles,cytometry may be used to determine the presence, quantity, and/ormodifications of specific proteins, nucleic acids, lipids,carbohydrates, or other molecules. Properties that may be measured bycytometry also include measures of cellular function or activity,including but not limited to phagocytosis, antigen presentation,cytokine secretion, changes in expression of internal and surfacemolecules, binding to other molecules or cells or substrates, activetransport of small molecules, mitosis or meiosis; protein translation,gene transcription, DNA replication, DNA repair, protein secretion,apoptosis, chemotaxis, mobility, adhesion, antioxidizing activity, RNAi,protein or nucleic acid degradation, drug responses, infectiousness, andthe activity of specific pathways or enzymes. Cytometry may also be usedto determine information about a population of cells, including but notlimited to cell counts, percent of total population, and variation inthe sample population for any of the characteristics described above.The assays described herein may be used to measure one or more of theabove characteristics for each cell, which may be advantageous todetermine correlations or other relationships between differentcharacteristics. The assays described herein may also be used toindependently measure multiple populations of cells, for example bylabeling a mixed cell population with antibodies specific for differentcell lines. A microscopy module may permit the performance of histology,pathology, and/or morphological analysis with the device, and alsofacilitates the evaluation of objects based on both physical andchemical characteristics. Tissues can be homogenized, washed, depositedon a cuvette or slide, dried, stained (such as with antibodies),incubated and then imaged. When combined with the data transmissiontechnologies described elsewhere herein, these innovations facilitatethe transmission of images from a CMOS/CDD or similar to a licensedpathologist for review, which is not possible with traditional devicesthat only perform flow cytometry. The cytometer can measure surfaceantigens as well as cell morphology; surface antigens enable moresensitive and specific tesing compared to traditional hematologylaboratory devices. The interpretation of cellular assays may beautomated by gating of one or more measurements; the gating thresholdsmay be set by an expert and/or learned based on statistical methods fromtraining data; gating rules can be specific for individual subjectsand/or populations of subjects.

In some embodiments, the incorporation of a cytometer module into apoint of service device provides the measurement of cellular attributestypically measured by common laboratory devices and laboratories forinterpretation and review by classically-trained medical personnel,improving the speed and/or quality of clinical decision-making. A pointof service device may, therefore, be configured for cytometric analysis.

Cytometric analysis may, for example, be by flow cytometry or bymicroscopy. Flow cytometry typically uses a mobile liquid medium thatsequentially carries individual cells to an optical, electrical oracoustic detector. Microscopy typically uses optical or acoustic meansto detect stationary cells, generally by recording at least onemagnified image. It should be understood that flow cytometry andmicroscopy are not entirely exclusive. As one example, flow cytometryassays may use microscopy to record images of cells passing by thedetector. Many of the targets, reagents, assays, and detection methodsmay be the same for flow cytometry and microscopy. As such, unlessotherwise specified, the descriptions below should be taken to apply tothese and other forms of cytometric analyses known in the art.

In some embodiments, a cytometry module may contain a microscopy stageand an objective. The microscopy stage may be configured to receive acytometry cuvette. The microscopy stage may be accessed by amodule-level sample handling system (e.g. configured to transport itemswithin a module) or a device-level sample handling system (e.g.configured to transport items between modules). A cytometry module maycontain a camera, CCD/CMOS sensor, or other imaging device operativelycoupled to the objective or microscopy stage, such that the imagingdevice may obtain a digital image of cells in a cuvette, assay unit, orother vessel. Cells in a cuvette, assay unit, or other vessel may besettled. The vessel may be fluidically isolated or independentlymovable. The digital image may be two-dimensional or three dimensional,and it may be a single image or a collection of images. The microscopicobjective can be finely positioned to focus the image via an actuator,such as by a cam connected to a motor. The objective may be focused onone or more planes of the sample. Focusing may be automated by imageanalysis procedures by computing the image sharpness of digital imagesamong other methods.

Flow Cytometry

Flow cytometry may be used to measure, for example, cell size (forwardscatter, conductivity), cell granularity (side scatter at variousangles), DNA content, dye staining, and quantitation of fluorescencefrom labeled markers. Flow cytometry may be used to perform cellcounting, such as by marking the sample with fluorescent markers andflowing past a sensing device. Cell counting may be performed onunlabeled samples as well.

Preferably up to 1000000 cells of any given type may be measured. Inother embodiments, various numbers of cells of any given type may bemeasured, including but not limited to more than or equal to about 10cells, 30 cells, 50 cells, 100 cells, 150 cells, 200 cells, 300 cells,500 cells, 700 cells, 1000 cells, 1500 cells, 2000 cells, 3000 cells,5000 cells, 6000 cells, 7000 cells, 8000 cells, 9000 cells, 10000 cells,100000 cells, 1000000 cells.

In some embodiments, flow cytometry may be performed in microfluidicchannels. Flow cytometry analysis may be performed in a single channelor in parallel in multiple channels. In some embodiments, flow cytometrymay sequentially or simultaneously measure multiple cellcharacteristics. Flow cytometry may be combined with cell sorting, wheredetection of cells that fulfill a specific set of characteristics arediverted from the flow stream and collected for storage, additionalanalysis, and/or processing. It should be noted that such sorting mayseparate out multiple populations of cells based on different sets ofcharacteristics, such as 3 or 4-way sorting.

Microscopy

Microscopy methods that may be used with this invention include but arenot limited to bright field, oblique illumination, dark field,dispersion staining, phase contrast, differential interference contrast(DIC), polarized light, epifluorescence, interference reflection,fluorescence, confocal (including CLASS), confocal laser scanningmicroscopy (CLSM), structured illumination, stimulated emissiondepletion, electron, scanning probe, infrared, laser, widefield, lightfield microscopy, lensless on-chip holographic microscopy, digital andconventional holographic microscopy, extended depth-of-field microscopy,optical scatter imaging microscopy, deconvolution microscopy, defocusingmicroscopy, quantitative phase microscopy, diffraction phase microscopy,confocal Raman microscopy, scanning acoustic microscopy and X-raymicroscopy. Magnification levels used by microscopy may include, asnonlimiting examples, up to 2×, 5×, 10×, 20×, 40×, 60×, 100×, 100×,1000×, or higher magnifications. Feasible magnification levels will varywith the type of microscopy used. For example, images produced by someforms of electron microscopy may involve magnification of up to hundredsof thousands of times. Multiple microscopy images may be recorded forthe same sample to generate time-resolved data, including videos.Individual or multiple cells may be imaged simultaneously, by parallelimaging or by recording one image that encompasses multiple cells. Amicroscopic objective may be immersed in media to change its opticalproperties, such as through oil immersion. A microscopic objective maybe moved relative to the sample by means of a rotating CAM to change thefocus. Cytometry data may be processed automatically or manually, andmay further include analyses of cell or tissue morphology, such as by apathologist for diagnostic purposes.

Cell counting can be performed using imaging and cytometry. Insituations where the subjects may be bright-field illuminated, thepreferred embodiment is to illuminate the subjects from the front with awhite light and to sense the cells with an imaging sensor. Subsequentdigital processing will count the cells. Where the cells are infrequentor are small, the preferred embodiment is to attach a specific ornon-specific fluorescent marker, and then illuminate the subject fieldwith a laser. Confocal scanning imaging is preferred. Preferably up to1000 cells of any given type may be counted. In other embodiments,various numbers of cells of any given type may be counted, including butnot limited to more than or equal to about 1 cell, 5 cells, 10 cells, 30cells, 50 cells, 100 cells, 150 cells, 200 cells, 300 cells, 500 cells,700 cells, 1000 cells, 1500 cells, 2000 cells, 3000 cells, 5000 cells.Cells may be counted using available counting algorithms. Cells can berecognized by their characteristic fluorescence, size and shape.

In some microscopy embodiments, brightfield illumination may be achievedby the use of a white light source along with a stage-condenser tocreate Koehler illumination. Brightfield images of cells, which maydetect properties similar to that of forward scattering in flowcytometry, can reveal cell size, phase-dense material within the cellsand colored features in the cell if the cells have been previouslystained. In one example embodiment, the Wright-Giemsa staining methodcan be used to stain human whole blood smear. Brightfield imaging showscharacteristic patterns of staining of human leukocytes. Thecharacteristically shaped red cells can also be identified in theseimages.

In some microscopy embodiments, darkfield imaging may be achieved by theuse of a ringlight based illumination scheme, or other epi- ortrans-darkfield illumination schemes available. Darkfield imaging may,for example, be used to determine light scattering properties of cells,equivalent to side scatter in flow cytometry, such as when imaging humanleukocytes. The internal and external features of the cell which scattermore light appear brighter and the features which scatter lesser amountsof light appear darker in a darkfield image. Cells such as granulocyteshave internal granules of size range (100-500 nm) which can scattersignificant amount of light and generally appear brighter in darkfieldimages. Furthermore, the outer boundary of any cell may scatter lightand may appear as a ring of bright light. The diameter of this ring maydirectly give the size of the cell. Microscopy methods may additionallybe used to measure cell volume. For example, red blood cell volume maybe measured. To increase accuracy, red blood cells may be transformedinto spheres through the use of anionic or zwitterionic surfactants, anddark field imaging used to measure the size of each sphere, from whichcell volumes may be calculated.

In some microscopy embodiments, small cells or formed elements which maybe below the diffraction-limited resolution limit of the microscope, maybe labeled with fluorescent markers; the sample may be excited withlight of appropriate wavelength and an image may be captured. Thediffraction pattern of the fluorescent light emitted by the labeled cellmay be quantified using computer analysis and correlated with the sizeof the cell. The computer programs used for these embodiments isdescribed elsewhere herein. To improve the accuracy of this method, thecells may be transformed into spheres by the use of anionic andzwitterionic surfactants.

Cell imaging may be used to extract one or more of the followinginformation for each cell (but is not limited to the following):

-   -   a. Cell size    -   b. Quantitative measure of cell granularity or light scattering        (also popularly called side scatter, based on flow cytometry        parlance)    -   c. Quantitative measure of fluorescence in each spectral channel        of imaging, after compensating for cross-talk between spectral        channels, or intracellular distribution pattern of fluorescent        or other staining    -   d. Shape of the cell, as quantified by standard and custom shape        attributes such as aspect ratio, Feret diameters, Kurtosis,        moment of inertia, circularity, solidity etc.    -   e. Color, color distribution and shape of the cell, in cases        where the cells have been stained with dyes (not attached to        antibodies or other types of receptor).    -   f. Intracellular patterns of staining or scattering, color or        fluorescence that are defined as quantitative metrics of a        biological feature such as morphology, for example density of        granules within cells in a darkfield image, or the number and        size of nucleolar lobes in a Giemsa-Wright stained image of        polymorphonuclear neutrophils etc.    -   g. Co-localization of features of the cell revealed in images        acquired in different channels.    -   h. Spatial location of individual cells, cellular structures,        populations of cells, intracellular proteins, ions,        carbohydrates and lipids or secretions (such as to determine the        source of secreted proteins).

A wide range of cell-based assays can be designed to use the informationgathered by cytometry. For example, an assay for performing a 5-partleukocyte differential may be provided. The reportables in this casemay, for example, be number of cells per microliter of blood for thefollowing types of leukocytes: monocytes, lymphocytes, neutrophils,basophils and eosinophils. Reportables may also be used to classifyleukocyte differentiation, or identify T and B-cell populations.

Fluorescence Microscopy

Fluorescence microscopy generally involves labeling of cells or othersamples with fluorescent labels, described in more detail below.Microscopic imaging of fluorescently labeled samples may gatherinformation regarding the presence, amounts, and locations of the targetthat is labeled at a given moment in time or over a period of time.Fluorescence may also be used to enhance sensitivity for detectingcells, cellular structures, or cellular function. In fluorescencemicroscopy, a beam of light is used to excite the fluorescent molecules,which then emit light of a different wavelength for detection. Sourcesof light for exciting fluorophores are well known in the art, includingbut not limited to xenon lamps, lasers, LEDs, and photodiodes. Detectorsinclude but are not limited to PMTs, CCDs, and cameras.

Electron Microscopy

Another nonlimiting example of microscopy uses electron beams instead ofvisible light, such as transmission electron microscopy (TEM) andscanning electron microscopy (SEM). In TEM, a beam of electrons istransmitted through a thin sample, and interactions between theelectrons and the specimens are mapped and magnified. TEM is thuscapable of imaging resolutions up to individual atoms. TEM contrast mayuse a bright field imaging mode, where electrons are absorbed by thesample; a diffraction contrast mode, where electrons are scattered bythe sample; electron energy loss spectroscopy (EELS), which detectselectrons that have interacted with specific components of a samplebased on their voltages; phase contrast or high-resolution transmissionelectron microscopy; diffraction, which produces characteristicdiffraction patterns that can be computationally analyzed to determinethe sample structure; three dimensional imaging, where the sample isrotated and imaged multiple times to reconstruct the overallthree-dimensional structure.

Samples for TEM may be prepared by forming a dilute solution ofmolecules or carving larger samples to a layer at most hundreds ofnanometers thick. For negative staining EM, biological samples aretypically spread on a grid, dried, and fixed with negative stainingreagents containing heavy metals, such as osmium, lead, uranium, orgold; one such staining reagent is uranyl acetate. For cryo-EM, samplesmay be embedded in vitreous ice and further cooled to liquid nitrogen orhelium temperatures.

In SEM, a focused electron beam is rastered over a surface to producesecondary electrons, back-scattered electrons, X-rays, light, current,and/or transmitted electrons. SEM can be used to visualize samples lessthan 1 nm in size with a large field depth to produce informationregarding the 3D surface structure of a sample. SEM using back-scatteredelectrons may be used with labels such as colloidal gold, for exampleattached to immunolabels, to better detect specific targets.

For SEM, samples typically contain no water. Biological samples such ascells may be fixated to preserve their internal structures beforedrying, such as by evaporation, heat, or with critical point drying,where water is sequentially replaced with an organic solvent, followedby liquid carbon dioxide. Conducting samples generally require little orno additional sample preparation, other than mounting onto a specimenholder compatible with the scanning electron microscope. Nonconductingsamples may be coated with a thin layer of a conducting material, suchas gold, gold/palladium, platinum, osmium, iridium, tungsten, chromium,or graphite, which may increase signal, increase resolution, anddecrease accumulation of static electric charges during irradiation.Other methods for increasing conductivity of an SEM sample includestaining with the OTO staining method. Nonconducting samples do notrequire increased conductivity for SEM imaging. As some nonlimitingexamples, environmental SEM and field emission gun (FEG) SEM may be usedto image nonconducting samples.

Reagents

Cells may be prepared for cytometry assays by any method known in theart. Cells may be optionally fixed, stained, and/or otherwise labeledwith a detectable marker. Cells may be fixed with a variety of methodsknown in the art, including but not limited to heat, freeze, perfusion,immersion, and chemical fixation. Chemical fixation may be achieved by awide variety of agents, including but not limited to crosslinking agents(such as formaldehyde, glutaraldehyde, other aldehydes, and theirderivatives), precipitating agents (such as ethanol and other alcohols),oxidizing agents (such as osmium tetroxide or potassium permanganate),potassium dichromate, chromic acid, mercury-containing fixatives, aceticacid, acetone, picrates, and HOPE fixative. Cells may also bepermeabilized, such as through the use of surfactants, as may be usefulfor subsequent internal labeling or staining.

Cells may be stained with any optically detectable dye, stains, orcoloring agents, such as nucleic acid dyes (including intercalatordyes), lipophilic dyes, protein dyes, carbohydrate dyes, heavy metalstains. Such dyes and stains or staining processes include but are notlimited to Acid Fast Bacilli staining, Alcian Blue staining, AlcianBlue/PAS staining, Alizarin Red, alkaline phosphatase staining,aminostyryl dyes, ammonium molybdate, Azure A, Azure B, BielschowskyStaining, Bismark brown, cadmium iodide, carbocyanines, carbohydrazide,carboindocyanines, Carmine, Coomassie blue, Congo Red, crystal violet,DAPI, ethidium bromide, Diff-Quik staining, eosin, ferric chloride,fluorescent dyes, fuchsin, Giemsa stain, Golgi staining, Golgi-Coxstaining, Gomori's Trichrome staining, Gordon Sweet's staining, Gramstaining, Grocott Methenamine staining, haematoxylin, hexamine, Hoechststains, Hyaluronidase Alcian Blue, indium trichloride,indocarbocyanines, indodicarbocyanines, iodine, Jenner's stain,lanthanum nitrate, lead acetate, lead citrate, lead(II) nitrate,Leishman stain, Luna staining, Luxol Fast Blue, Malachite green, MassonFontana staining, Masson Trichrome staining, methenamine, methyl green,methyline blue, microglia staining, Miller's Elastic staining, neutralred, Nile blue, Nile red, Nissl staining, Orange G, osmium tetroxide,Papanicolaou staining, PAS staining, PAS diastase staining, periodicacid, Perls Prussian Blue, phosphomolybdic acid, phosphotungstic acid,potassium ferricyanide, potassium ferrocyanide, Pouchet staining,propidium iodide (PI), Prussian Blue, Renal Alcian Blue/PAS staining,Renal Masson Trichrome staining, Renal PAS Methenamine staining,Rhodamine, Romanovsky stain, Ruthenium Red, Safranin O, silver nitrate,Silver staining, Sirius Red, sodium chloroaurate, Southgate'sMucicannine, Sudan staining, Sybr Green, Sybr Gold, SYTO dyes, SYPROstains, thallium nitrate, thiosemicarbazide, Toluidine Blue, uranylacetate, uranyl nitrate, van Gieson staining, vanadyl sulfate, von Kossastaining, WG staining, Wright-Giemsa stain, Wright's stain, X-Gal, andZiehl Neelsen staining Cells may be treated with uncolored dyeprecursors that are converted to a detectable product after treatment,such as by enzymatic modification (such as by peroxidases orluciferases) or binding to an ion (such as Fe ions, Ca²⁺ or H⁺).

Cells may further be labeled with fluorescent markers. Usefulfluorescent markers include natural and artificial fluorescentmolecules, including fluorescent proteins, fluorophores, quantum dots,and others. Some examples of fluorescent markers that may be usedinclude but are not limited to: 1,5 IAEDANS; 1,8-ANS;5-carboxy-2,7-dichlorofluorescein; 5-Carboxyfluorescein (5-FAM);fluorescein amidite (FAM); 5-Carboxynapthofluorescein;tetrachloro-6-carboxyfluorescein (TET); hexachloro-6-carboxyfluorescein(HEX); 2,7-dimethoxy-4,5-dichloro-6-carboxyfluorescein (JOE); VIC®;NED™; tetramethylrhodamine (TMR); 5-Carboxytetramethylrhodamine(5-TAMRA); 5-HAT (Hydroxy Tryptamine); 5-Hydroxy Tryptamine (HAT); 5-ROX(carboxy-X-rhodamine); 6-Carboxyrhodamine 6G; 6-JOE; Light Cycler® red610; Light Cycler® red 640; Light Cycler® red 670; Light Cycler® red705; 7-Amino-4-methylcoumarin; 7-Aminoactinomycin D (7-AAD);7-Hydroxy-4-methylcoumarin; 9-Amino-6-chloro-2-methoxyacridine; ABQ;Acid Fuchsin; ACMA (9-Amino-6-chloro-2-methoxyacridine); AcridineOrange; Acridine Red; Acridine Yellow; Acriflavin; Acriflavin FeulgenSITSA; AutoFluorescent Proteins; Texas Red and related molecules;Thiadicarbocyanine (DiSC3); Thiazine Red R; Thiazole Orange; Thioflavinderivatives; Thiolyte; Thiozole Orange; Tinopol CBS (Calcofluor White);TMR; TO-PRO-1; TO-PRO-3; TO-PRO-5; TOTO-1; TOTO-3; TriColor (PE-Cy5);TRITC (TetramethylRodamine-lsoThioCyanate); True Blue; TruRed;Ultralite; Uranine B; Uvitex SFC; WW 781; X-Rhodamine; XRITC; XyleneOrange; Y66F; Y66H; Y66W; YO-PRO-1; YO-PRO-3; YOYO-1; interchelatingdyes such as YOYO-3, Sybr Green, Thiazole orange; members of the AlexaFluor® dye series (from Molecular Probes/Invitrogen) such as Alexa Fluor350, Alexa Fluor 405, 430, 488, 500, 514, 532, 546, 555, 568, 594, 610,633, 635, 647, 660, 680, 700, and 750; members of the Cy Dye fluorophoreseries (GE Healthcare), such as Cy3, Cy3B, Cy3.5, Cy5, Cy5.5, Cy7;members of the Oyster® dye fluorophores (Denovo Biolabels) such asOyster-500, -550, -556, 645, 650, 656; members of the DY-Labels series(Dyomics), such as DY-415, -495, -505, -547, -548, -549, -550, -554,-555, -556, -560, -590, -610, -615, -630, -631, -632, -633, -634, -635,-636, -647, -648, -649, -650, -651, -652, -675, -676, -677, -680, -681,-682, -700, -701, -730, -731, -732, -734, -750, -751, -752, -776, -780,-781, -782, -831, -480XL, -481XL, -485XL, -510XL, -520XL, -521XL;members of the ATTO series of fluorescent labels (ATTO-TEC GmbH) such asATTO 390, 425, 465, 488, 495, 520, 532, 550, 565, 590, 594, 610, 611X,620, 633, 635, 637, 647, 647N, 655, 680, 700, 725, 740; members of theCAL Fluor® series or Quasar® series of dyes (Biosearch Technologies)such as CAL Fluor® Gold 540, CAL Fluor® Orange 560, Quasar® 570, CALFluor® Red 590, CAL Fluor® Red 610, CAL Fluor® Red 635, Quasar® 570, andQuasar® 670.

Fluorescent markers may be coupled to a targeting moiety to allowspecific binding or localization, for example, to a specific populationof cells, of which there are many known in the art. Nonlimiting examplesinclude antibodies, antibody fragments, antibody derivatives, aptamers,oligopeptides such as the nuclear localization sequence (NLS), smallmolecules that serve as specific ligands for receptors including manyhormones and drugs, nucleic acid sequences (such as for FISH), nucleicacid binding proteins (including repressors and transcription factors),cytokines, ligands specific for cellular membranes, enzymes, moleculesthat specifically bind to enzymes (such as inhibitors), lipids, fattyacids, and members of specific binding interactions such asbiotin/iminobiotin and avidin/streptavidin.

Targets for specific labeling may be natural or artificial and mayencompass proteins, nucleic acids, lipids, carbohydrates, smallmolecules, and any combinations thereof. These include intracellular andcell surface markers. Intracellular markers include any molecule,complex, or other structure within the cell. A few nonlimiting examplesinclude genes, centromeres, telomeres, nuclear pore complexes,ribosomes, proteasomes, an internal lipid membrane, metabolites such asATP, NADPH, and their derivatives, enzymes or enzyme complexes, proteinchaperones, post-translational modifications such as phosphorylation orubiquitinylation, microtubules, actin filaments, and many others. Cellsurface markers include but are not limited to proteins such as CD4,CD8, CD45, CD2, CRTH2, CD19, CD3, CD14, CD36, CD56, CD5, CD7, CD9, CD10,CD11b, CD11c, CD13, CD15, CD16, CD20, CD21, CD22, CD23, CD24, CD25,CD33, CD34, CD37, CD38, CD41, CD42, CD57, CD122, CD52, CD60, CD61, CD71,CD79a, CD95, CD103, CD117, CD154, GPA, HLA, KOR, FMC7. In someembodiments, the targets may be specific regions within a cell, such astargeting to the interior of specific organelles or membrane-boundvesicles. In some embodiments, the target may be the result of geneticor other manipulation, such as cloning Lac binding sites into a geneticsequence for targeted binding by a labeled Lac protein.

Cells may be labeled through various means, including but not limited tosurface labeling, permeabilization of the cell membrane and/or cellwall, active transport or other cellular processes, diffusion throughthe membrane, carrier particles such as lipid vesicles or otherhydrophobic molecules, and production by the cell (such as forrecombinantly fluorescent proteins).

In some embodiments, samples containing mixed populations of cells maybe treated before optical detection to enrich for detection of targetpopulation(s) of cells. Some example methods for enrichment include butare not limited to centrifugation, sorting (with or without labeling),selective killing of non-target cells such as by lysis, and selectivelabeling to improve detection of target cells. For imaging, cells may besuspended in liquid medium (as is preferred for flow cytometry),attached to a surface, or confined in a small volume, such as in amicrofluidic well or channel.

One or more agents such as cell activators, stimulators, or inhibitors,may be added to the entire sample, or portions of the sample, todetermine how the cells/samples respond. Such agents can be non-specific(such as cytokines), or specific (such as antigens), or a combinationthereof. Tissue samples may be cultured in the presence of one or moreagents for different periods of time under different environmentalconditions and analyzed in real time. Culture conditions can be variedover time based on measured response, and additional agents added overtime as required. Also, one may examine sensitivity to certain drugs,such as resistance to antibiotics, using these techniques. The samplesmay be analyzed before, during and after agent administration. Exposurewith one or more agents can be sequential and/or repeated over time. Theconcentration of the agents can be titrated based on measured responses.

Tissue samples (such as from biopsy) may be homogenized in a variety ofways, including through the use of a grinder, a pulverizer, actuation bypipette/nozzles, or centrifugation with or without beads (such as nanosharp beads), pushing the sample through a mesh and/or micro-column, orultrasonication. Fluorescence activated cell sorting (FACS) may beperformed with the inclusion of flow and/or other cell-separationmethods (such as magnetic separation).

Spectroscopy

Spectroscopy includes any and all assays that produce luminescence orchange light (e.g., color chemistry). These may include one or more ofthe following: spectrophotometry, fluorimetry, luminometry,turbidimetry, nephelometry, refractometry, polarimetry, and measurementof agglutination.

Spectrophotometry refers to measuring a subject's reflection ortransmission of electromagnetic waves, including visible, UV, andinfrared light. Spectrophotometry may, for example, be used to determinenucleic acid concentrations in a sample, such as by measuring absorbanceat a wavelength of about 260, to determine protein concentration bymeasuring absorbance at a wavelength of about 280, and/or to determinesalt concentration by measuring absorbance at a wavelength of about 230.

Other examples of spectrophotometry may include infrared (IR)spectroscopy. Examples of infrared spectroscopy include near-infraredspectroscopy, far-infrared spectroscopy laser-Raman spectroscopy, Ramanconfocal laser spectroscopy, Fourier Transform infrared spectroscopy,and any other infrared spectroscopy technique. Frequencies of less thanabout 650 cm-1 are typically used for far-infrared spectroscopy,frequencies greater than about 4000 cm-1 are typically used fornear-infrared spectroscopy, while frequencies between about 650 andabout 4000 cm-1 are typically used for other types of IR spectroscopy.IR spectroscopy has many biomedical applications, including in cancerdiagnosis, arthritis diagnosis, determining chemical compositions ofbiological fluids, determining septic state, and others. IR spectroscopymay be used on solid samples, such as tissue biopsies, cell cultures, orPap smears; or on liquid samples, such as blood, urine, synovial fluid,mucus, and others. IR spectroscopy may be used to differentiate betweennormal and cancerous cells as described in U.S. Pat. No. 5,186,162,herein incorporated by reference. IR spectroscopy may also be used onblood samples to detect markers for cancers of various solid organs. IRspectroscopy may also be used to determine cellular immunity inpatients, such as to diagnose immunodeficiencies, autoimmune disorders,infectious diseases, allergies, hypersensitivity, and tissue transplantcompatibility.

IR spectroscopy may be used to determine glucose levels in blood, whichis of use for diabetic patients, such as for monitoring insulinresponse. IR spectroscopy may further be used to measure othersubstances in blood samples, such as alcohol levels, fatty acid content,cholesterol levels, hemoglobin concentration. IR spectroscopy can alsodistinguish between synovial fluid from healthy and arthritic patients.

Fluorimetry refers to measuring the light emitted by a fluorescentmolecule coupled to a subject upon exciting the fluorescent moleculewith incident light. Fluorimetry may use any of the fluorescentmolecules, labels, and targets as described for cytometric assays above.In some embodiments, fluorimetry uses substrate molecules that change influorescence based on an enzymatic activity, such as converting NAD+ toNADH or vice versa or producing beta-galactosidase from a precursormolecule. Fluorimetry may be used with a polarized excitation source tomeasure fluorescence polarization or anisotropy of a subject, which mayprovide information about the size and/or binding state.

Colorimetry refers to measuring the transmissive color absorption of asubject, preferably by backlighting the subject with white light withthe result sensed by an imaging sensor. Examples include some assaysthat use oxidases or peroxidases combined with a dye that becomescolored in the presence of hydrogen peroxide. One method that measuresperoxidase activity in whole cell suspensions of human white blood cellsis disclosed in Menegazzi, et al., J. Leukocyte Biol 52: 619-624 (1992),which is herein incorporated by reference in its entirety. Such assaysmay be used to detect analytes that include but are not limited toalcohols, cholesterols, lactate, uric acid, glycerol, triglycerides,glutamate, glucose, choline, NADH. Some of the enzymes that may be usedinclude horseradish peroxidase, lactoperoxidase, microperoxidase,alcohol oxidase, cholesterol oxidase, NADH oxidase. Other nonlimitingexamples of colorimetric assays include dye-based assays to determineprotein concentration, such as Bradford, Lowry, biureat, and Nano-orangemethods. The pH of a sample may also be determined by colorimetricassays with indicator dyes, including but not limited to phenolphtalein,thymolphtalein, alizarin Yellow R, indigo carmine, m-cresol purple,cresol red, thymol blue, xylenol blue,2,2′,2″,4,4′-pentamethoxytriphenyl carbinol, benzopurpurin 4B, metanilyellow, 4-phenylazodiphenylamine, malachite green, quinaldine red,orange IV, thymol blue, xylenol blue, and combinations thereof.

Luminometry uses no illumination method as the subject emits its ownphotons. The emitted light can be weak and can be detecting using anextremely sensitive sensor such as a photomultiplier tube (PMT).Luminometry includes assays that produce chemiluminescence, such asthose using luciferases or some assays using peroxidases.

In some embodiments, systems, devices, methods, or assays providedherein include a chemiluminescent compound. In some embodiments,chemiluminescent compounds may emit light, such as upon a chemicalalteration (e.g. oxidation, phosphorylation, dephosphorylation,hydrolysis, etc.) of the original chemiluminescent compound.Chemiluminescent compounds may include, for example:3-(2′-spiroadamantyl)-4-methoxy-4-(3″-phosphoryloxy)-phenyl-1,2-dioxetane(AMPPD), luminol, N-(4-aminobutyl)-N-ethylisoluminol,4-aminophthalhydrazide, coelenterazine hcp, coelenterazine fcp, andD-luciferin. These or other chemiluminescent compounds may be provided,for example, in an assay unit, reagent unit, vessel, tip, or containerin a cartridge or assay station provided herein and may be used insystems configured for discretely multiplexing assays with other ofother assay methodologies (that may be the same or different).Chemiluminescent compounds may be provided in various forms, including,for example, in lyophilized, gel, or liquid forms. In some embodiments,a chemiluminescent enhancer molecule (for example,4-(4,5-diphenyl-2-imidazolyl) phenol) is provided with achemiluminescent compound.

For turbidimetry, the preferred embodiment for sensing is backlightingthe subject with white light with the result sensed by an imagingsensor. For turbidimetry, the reduction of the intensity of thetransmitted light is measured. Turbidimetry may be used, for example, todetermine a concentration of cells in solution. In some embodiments,turbimetry is measured by nephelometry.

Nephelometry measures the light that is transmitted or scattered afterpassing through a subject in a suspension. In some embodiments, thesubject in suspension is a substrate bound to an immunoglobin such asIgM, IgG, and IgA, or salts which have precipitated out of solution

Polarimetry measures the polarization of, typically, electromagneticwaves by a subject. Polarimetry assays include circular dichroism, whichmay provide structural information and light scattering assays, whichmay provide information about the size and/or shape of the subject. Onenonlimiting example of light scattering assays uses dynamic lightscattering (DLS). Subjects for these assays do not require labeling.

Chromogens

In some embodiments, systems, devices, methods, or assays (including,for example, colorimetric assays, absorbance assays, fluorescent assays,and turbimetric assays) provided herein include a chromogen (also termedherein, e.g., colorants, colored products, and other terms). In someembodiments, chromogens may be capable of conversion from a first colorto a second color, such as upon a chemical alteration (e.g. oxidation,phosphorylation, dephosphorylation, etc.) of the original chromogenmolecule. In some instances, a chromogen is an essentially colorlessmolecule which converts into a colored pigment upon chemical alterationof the molecule. Formation of chemically altered chromogen product maybe monitored, for example, by observing a decrease in the level of theoriginal, non-chemically altered chromogen, or by observing an increasein the level of the chemically altered chromogen. Levels of a particularchromogen (chemically altered or non-chemically altered) may bemonitored, for example, by measuring the absorbance of one or moreselected wavelength(s) of light by a sample which may contain thechromogen. For such measurements, commonly, the monitored wavelength(s)is a wavelength of light that the chromogen absorbs. In such instances,higher amounts of the chromogen in a sample are correlated with higherabsorbance of the selected wavelength of light by the sample. Thesechromogen(s) may be used in systems configured for discretelymultiplexing assays with other of other assay methodologies (that may bethe same or different).

Chromogens that may be used with systems, devices, and methods providedherein may include, for example, i) substrates which may be oxidized(e.g. molecules that change color upon oxidization, such as byperoxidase and hydrogen peroxide), for example: aniline and relatedderivatives [e.g. 2-amino-4-hydroxybenzenesulfonic acid (AHBS)(forms ayellow dye upon oxidation which may be monitored at 415 nm),N-(2-hydroxy-3-sulfopropyl)-3,5-dimethyoxyaniline (ALPS) coupled withAAP (forms a dye upon oxidation that may be monitored at 610 nm), N, Ndiethylaniline], o-dianisidine (forms a yellow-orange dye upon oxidationthat may be monitored at 405 nm), 10-acetyl-3,7-dihydroxyphenoxazine(ADHP) (forms a dye upon oxidation that may be monitored, for example,colorimetrically at 570 nm or fluorescently at EX/EM=535/587 nm)resazurin (7-hydroxy-3H-phenoxazin-3-one 10-oxide) and relatedderivatives [e.g 10-acetyl-3,7-dihydroxyphenoxazine (Amplex Red) and7-ethoxyresorufin (form a pink color on oxidation which may be monitoredcolorimetrically or fluorescently at EX/EM=570/585); ii) substrates ofphosphatases (e.g. molecules that change color upon dephosphorylation),for example: p-nitrophenyl phosphate (pNPP) (forms p-nitrophenol upondephosphorylation, which may be measured by absorbance at 405 nm); iii)substrates of hydrolases (e.g. molecules that change color uponhydrolysis), for example: ortho-nitrophenyl-beta-galactoside (ONPG) (maybe hydrolyzed by beta-galactosidase to galactose and ortho-nitrophenol;ortho-nitrophenol may be measured by absorbance at 420 nm); iv)substrates which may change color upon complex formation, for example:o-cresolphthalein (forms a complex with calcium, which may be measuredby absorbance at 575 nm), potassium cyanide (forms a complex withhemoglobin, which may be measured by absorbance at 540 nm), thiocyanate(forms a complex with iron, which may be measured by absorbance at 480nm), 2,4,6-Tripyridyl-s-triazine (TPTZ) (forms a complex with iron,which may be measured by absorbance at 620 nm); v) substrates which maybe insoluble upon complex formation, for example: tetraphenylborate(forms a complex with potassium, which may precipitate out of solution);vi) substrates which may change color upon a change in pH (pHindicators), for example: bromophenol blue, methyl red, litmus,phenolphthalein and phenol red. These or other chromogens may beprovided, for example, in an assay unit, reagent unit, vessel, tip, orcontainer in a cartridge, assay station, or device provided herein.Chromogens may be provided in various forms, including, for example, inlyophilized, gel, or liquid forms.

Radioactivity Assays

Radioactive assays use at least one radioactive isotype as a detectablelabel. Radioactive labels may be used as labels for imaging or tocalculate enzymatic activity. Such enzymatic assays may be measured atthe end of the reaction (endpoint assays) or measured multiple timesover the course of the reaction (time course assays). As a nonlimitingexample, ATP labeled with ³²P on the gamma phosphate may be used toassay activity of ATPases present in the sample. In another embodiment,a labeled precursor compound or other molecule may be introduced to acell or other sample to measure synthesis of a product molecule (a“pulse”). Such introduction of a labeled precursor may be followed byaddition of an unlabeled version of the precursor (a “chase”). Someexamples of pulse-chase assays include but are not limited to using³H-leucine as a precursor for insulin synthesis and ³⁵S-methionine as aprecursor for protein synthesis. It should be noted that these types ofassays do not necessarily require the use of a radioactive label, as isknown to one familiar in the art.

Mass Spectrometry

In some embodiments, at least a portion of the sample may be analyzed bymass spectrometry. The sample may be provided to the mass spectrometeras a solid, liquid, or gas, and any of a variety of ionizationtechniques may be used, including matrix-assisted laserdesorption/ionization (MALDI), electrospray (including electrospray,microspray, and nanospray), inductively coupled plasma (ICP), glowdischarge, field desorption, fast atom bombardment, thermospray,desorption/ionization on silicon, atmospheric pressure chemicalionization, DART, secondary ion mass spectrometry, spark ionization,thermal ionization, and ion attachment ionization. Ionization may formpositive or negative ions. Methods for performing these techniques arewell-known in the art.

For solid and liquid phase mass spectrometry, samples may be presentedon a sample presentation apparatus composed of any suitable material,which may be solid or liquid. The sample presentation surface may haveattached enzymes or enzyme complexes that chemically modify or bind tothe sample. Examples of chemical modification include but are notlimited to enzymatic cleavage, purification, and adding a chemicalmoiety.

In MALDI, samples are typically premixed with a highly absorbing matrix,then bombarded with laser light for ionization. Samples for MALDI aretypically thermolabile, non-volatile organic compounds of high molecularmass, preferably up to 30,000 Da. Samples may be presented in anyappropriate volatile solvent. For positive ionization, trace amounts oftrifluoroacetic acid may be used. The MALDI matrix may be any materialthat solubilizes biomolecules, absorbs light energy at a frequencyeasily accessible by a laser, and is unreactive with respect tobiomolecules. Suitable matrices include nicotinic acid, pyrozinoic acid,vanillic acid, succinic acid, caffeic acid, glycerol, urea or trisbuffer (pH 7.3). Preferable matrices include a-cyano-4-hydroxycinnamicacid, ferulic acid, 2,5-dihydroxybenzoic acid, sinapic (or sinapinic)acid, 3,5-dimethoxy, 4-hydroxy-trans-cinnamic acid, other cinnamic acidderivatives, gentisic acid and combinations thereof.

In electrospray ionization (ESI), samples are typically dissolved in avolatile polar solvent, such as an acetonitrile solution, andaerosolized by a strong voltage (for example, 3-4 kV, or lower forsmaller samples, such as are used in microspray and nanospray) at acapillary tip. Samples for ESI typically range from less than 100 Da tomore than 1 Mda in mass. Aerosolization may be enhanced by flowing anebulizing gas past the capillary tip, such as nitrogen gas. Theresulting charged droplets are further decreased in size by solventevaporation, aided by a drying gas such as nitrogen that is typicallyheated. Additional reagents may be added to the solvent to aid inionization. As nonlimiting examples, trace amounts of formic acid mayaid protonation of the sample for positive ionization, while traceamounts of ammonia or a volatile amine may aid deprotonation of thesample for negative ionization.

Analytes for mass spectrometry include but are not limited to proteins,carbohydrates, lipids, small molecules, and modifications and/orcombinations thereof. Usually, proteins and peptides are analyzed withpositive ionization, while saccharides and oligonucleotides are analyzedwith negative ionization. Analytes may be analyzed whole or infragments. Mass spectrometry may be used to determine the composition ofa mixture, total size of subject(s), chemical structures, andsequencing, such as of oligopeptides or oligonucleotides. In someembodiments, mass spectrometry can be used to determine bindinginteractions, such as (but not limited to) between protein and ligandsincluding small molecules, peptides, metal ions, nucleic acids, andother small molecules.

In some embodiments, tandem mass spectrometry may be used, where two ormore analyzers are used in sequence, separated by a collision cell tofragment the subject ions. Tandem MS thus is capable of firstdetermining the overall mass of a subject, followed by determiningadditional structural information based on how the subject fragments.Examples of tandem spectrometry include, but are not limited toquadrupole—quadrupole, magnetic sector—quadrupole, magneticsector—magnetic sector, quadrupole—time-of-flight. Tandem spectrometryis particularly suited for determining structures, including of smallorganic molecules and for peptide or oligonucleotide sequencing. Duallight source for measuring absorbance and/or fluorescence, comprising ofa broad-band light source for absorbance measurement and a laser diodefor fluorescence measurement. CCD-based compact spectrophotometerstypically use an FPGA/CLPD to control acquisition; however,spectrometers provided herein use a general purpose microprocessor,which may offer more flexibility in terms of general-purpose computing,as well as the ability to update firmware remotely. In addition, thespectrometer can be equipped with a general purpose camera which enablesinterrogation of the sample before a reading to ensure sample/vesselintegrity. Feedback such as this helps in reducing catastrophicfailures, and allows for real-time correction. At least some embodimentsof systems herein may have one or more stations that include a massspectrometry station to configured to receive individual samplevessel(s) or arrays of sample vessels.

X-Ray Photoelectron Spectroscopy

X-ray Photoelectron Spectroscopy (XPS) or Electron Spectroscopy forChemical Analysis (ESCA) is a photoelectron spectroscopic analysismethod for detecting photoelectrons emitted by surfaces of samples todetermine their composition. Photoelectron spectroscopic analysis may befurther classified according to light source as XPS and UV photoelectronspectroscopy (UPS).

ESCA involves irradiating a sample surface with ultraviolet or x-raysand detecting the characteristic photoelectrons emitted by the elementsof the sample. XPS specifically refers to ESCA using x-rays. Thephotoelectrons are filtered by an electrostatic or magnetic analyzerwhich allows only electrons of a specified narrow energy band to passthrough to a detector. The binding energy of the emitted electrons isunique for each element, allowing identification of each element on thesurface. The intensity of the detected beam typically represents theconcentration of a given chemical constituent on or near a specimensurface. U.S. Pat. No. 3,766,381, herein incorporated by reference,describes such a system. ESCA and XPS may detect any element with anatomic number of 3 or above, and may detect the compositions of samplesup to l Onm from the surface. As a result, ESCA and XPS are particularlysuited to determine empirical formulas of pure materials, to detectcontaminants as low as parts per million, and to detect the chemical orelectronic state of each element of a sample surface. In XPS, theemitted electrons typically have short inelastic free paths in solidsamples. As a result, further information about the amount of an element(such as the depth an element extends from the surface) may bedetermined by analyzing the angle at once the emitted electrons emergefrom the surface. ESCA/XPS may be used to analyze samples including butnot limited to inorganic compounds, semiconductors, polymers, metalalloys, elements, catalysts, glasses, ceramics, paints, papers, inks,woods, plant parts, make-up, teeth, bones, medical implants,bio-materials, viscous oils, glues, ion-modified materials.

Another method of sample analysis uses Auger electrons, called Augerelectron spectroscopy (AES), which functions similarly to ESCA, exceptthat it uses a beam of electrons instead of UV or X-rays.

Chromatography

Chromatography methods use different properties of solutes in a mixtureto allow separation. Many different chromatography methods are known inthe art, including but not limited to paper chromatography, thin layerchromatography (TLC), column chromatography gas chromatography, liquidchromatography, affinity chromatography, displacement chromatography,ion exchange chromatography (cation and anion), hydrophobic interactionchromatography, size exclusion chromatography such as gel filtration,perfusion chromatography, push column chromatography, reversed-phasechromatography, two-dimensional chromatography, high performance liquidchromatography, packed capillary chromatography, open tubular liquidchromatography, pyrolysis gas chromatography, chiral chromatography, andmany others.

Chromotography typically relies on a solid stationary phase and a mobilephase (a solvent) that carries the sample. The stationary phase cancomprise a solid polymer, e.g., plastic, glass, other polymers, paper,cellulose, agarose, starch, sugars, magnesium silicate, calcium sulfate,silicic acid, silica gel, florisil, magnesium oxide, aluminum oxide(alumina), activated charcoal, diatomaceous earth, perlite, clays, orother similar substances known in the art. The stationary phase may betreated or otherwise modified to have a characteristic that slows themobility of at least one solute in the sample mixture. For ion exchangechromatography, the stationary phase may comprise a charged residue, forexample an anion that attracts positively charged solutes. For sizeexclusion chromatography, the stationary phase may comprise pores,tunnels, or other structures that may slow migration of smaller solutescompared to larger solutes. For affinity chromatography, the stationaryphase may comprise a binding moiety that specifically recognizes somesolutes. Typically, different solutes have different distributionequilibria. Therefore, different solutes will move across the stationaryphase at differing rates depending on their relative affinity for thestationary phase on one hand and for the solvent on the other. As thecomponents of the mixture (i.e., analytes) are separated, they begin toform moving bands or zones, which may be detected on the stationaryphase, as is typical for example on TLC, or as they are sequentiallyeluted, as is typical but not required for column chromatographymethods.

Separation results depend on many factors, including, but not limitedto, the stationary phase chosen, polarity of the solvent, size of thestationary phase (such as length and diameter of columns) relative tothe amount of material to be separated, and the rate of elution. In somecases, a long column or multiple columns arranged in series may berequired to separate the sample effectively. This is particularly truewhen the sample has a relatively low distribution equilibrium betweenthe stationary phase and the solvent. In other cases, the sample canbind tightly to the adsorbent material and may require a differentsolvent to elute the sample from the adsorbent. As one nonlimitingexample, proteins or peptides with molecular weight of greater than 1000in aqueous medium bind tightly to a C-18 alkyl stationary phase. Thisbonding is so strong that it is difficult to effectively remove theprotein from the stationary phase using water. Typically an organiceluent, such as acetonitrile, alcohol (e.g., methanol, ethanol, orisopropanol), other relatively polar organic solvents (e.g., DMF), ormixtures thereof, may be used as an eluent to remove the protein fromthe stationary phase. Other examples include binding chromatographycolumns where the sample binds the stationary phase with such highaffinity that a competing binder is required to elute the sample.

Chromatography methods may be used to separate nearly any substance froma mixture. A few nonlimiting examples include separating specifichormones, cytokines, proteins, sugars, or small molecules such as drugsfrom biological samples such as blood. The separated samples may bedetected more easily after elution, or may be subjected to furtherseparation, purification, or processing. For example, nucleic acids maybe separated from a sample and used as templates for nucleic acidamplification. Other samples may also be separated, such as separatingtoxins from environmental samples or targets of interest from lysedcells.

Ion Exchange Chromatography

Ion exchange chromatography relies on charge-charge interactions betweenthe components of a sample and charges on the stationary phase (such asresin packed in a column) and/or mobile phase. In cation exchangechromatography, positively charged solutes bind to negatively chargedstationary phase molecules, while in anion exchange, negatively chargedsolutes bind to positively charged stationary phases. In typicalembodiments, the solutes bind to the column in a solvent of low ionicstrength, then the bound molecules are eluted off using an increasinggradient of a second elution solvent with a higher ionic strength. Insome examples, the gradient changes the pH or salt concentrations of theeluent solvent. Ion exchange is well suited for purifying nucleic acids,which are typically negatively charged, from mixed samples.

Common resins for anion exchange chromatography include but are notlimited to Q-resins, and diethylaminoethane (DEAE) resin. Cationexchange resins include but are not limited to S resins and CM resins.Some commercially available resins include Nuvia, UNOsphere, AG,Bio-Rex, Chelex, Macro-Prep MonoBeads, MiniBeads, Resource Q, SourceMedia, Capto IEX, Capto MMC, HiScreen IEX, HiPrep IEX, Sepharose FastFlow, HiLoad IEX, Mono Q, Mono S, and MacroCap SP. Buffers for anionexchange include but are not limited to N-methyl piperazine, piperazine,L-histidine, bis-Tris, bis-Tris propane, triethanolamine, Tris,N-methyl-diethanolamine, diethanolamine, 1,3-diaminopropane,ethanolamine, piperazine, 1,3-diaminopropane, piperidine, and phosphatebuffer. Buffers for cation exchange include maleic acid, malonic acid,citric acid, lactic acid, formic acid, butaneandioic acid, acetic acid,malonic acid, phosphate buffer, HEPES buffer, and BICINE.

Size-Exclusion Chromatography

Size-exclusion chromatography (SEC) separates solutes based on theirsize, and is typically used for large molecules or macromolecularcomplexes. In SEC, the stationary phase consists of porous particlessuch that molecules smaller than the pore size may enter the particles.As a result, smaller solutes have a longer flow path and a longertransit time through the SEC column and are separated from largersolutes that cannot fit in the pores. Size-exclusion chromatography mayuse aqueous or organic solvents, which may be known as gel-filtration orgel permeation chromatography, respectively. Size-exclusionchromatography may also be used to determine general size informationabout the solutes when compared to a standard macromolecule of knownsize. Size-exclusion chromatography is also affected by the shape of thesolute, such that exact size determinations typically cannot be made. Inone example, size-exclusion chromatography may be combined with dynamiclight scattering to obtain absolute size information on proteins andmacromolecules. Resins for SEC may be selected based on the size of thetarget solute to increase separation on the chromatography column.Commercially available resins for size-exclusion chromatography includeSuperdex, Sepharcryl, Sepharose, and Sephadex resins. Examples ofbuffers for SEC include but are not limited to Tris-NaCl, phosphatebuffered saline, and Tris-NaCl-urea.

Affinity Chromatography

Affinity chromatography uses differences in affinities of individualsolutes for a surface such as by chelation, immunochemical bonding,receptor-target interactions, and combinations of these effects. Anysample for which a suitable binding partner is known, preferably with adissociation constant (K_(d)) of 10⁻⁶ or less, may be separated byaffinity chromatography. In some embodiments, the target may beengineered to contain an artificial binding moiety, such as apoly-Histidine, polyarginine, polylysine, GST, MBP, or other peptide tag(which may be removed subsequent to chromatography). Ligands and theirtarget molecules for affinity chromatography include but are not limitedto biotin and avidin and related molecules, monoclonal or polyclonalantibodies and their antigens, procainamide and cholinesterase, N-methylacridinium and acetylcholinesterase; P-aminobenzamidine and trypsin;P-aminophenol-beta-D-thiogalacto-pyranoside and beta-galactosidase;chitin and lysozyme; methotrexate and dihydrofolate reductase; AND andalcohol dehydrogenase; sulfanilamide and carbonic anhydrase; DNA and DNApolymerase; complementary nucleic acid sequences; oxidized glutathioneand glutathione reductase; P-aminobenzamidine and urokinase; trypsin andsoybean trypsin inhibitor; N 6-aminocaproyl-3′,5′-cAMP and ProteinKinase; Pepstatin and Renin; 4-Chlorobenzylamine and Thrombin;N-(4-amino phenyl) Oxamic Acid and Influenza Virus; Prealbumin andRetinal-binding Protein; Neurophysin and Vasopressin; Lysine andPlasminogen; Heparin and Antithrombin; Cycloheptaamylose and Human SerumAmylase; Cortisol and Transcortin; Pyridoxal-5-phosphate andGlutamate-pyruvate transaminase; Chelating Agents and Metal Ions;Chelating Agent-Cu and Superoxide Dismutase; Chelating Agent-Zn andHuman Fibrinogen; Coenzyme A and Succinic Thiokinase; Flavin andLuciferase; Pyridoxal Phosphate and Tyrosine Aminotransferase; Porphyrinand Haemopexin; Lysine and Ribosomal RNA; Polyuridine and mRNA;Concanavalin A and Immunoglobulins; 3-phospho-3-hydroxypropionate andEnolase; D-malate and Fumarate Hydratase; Atropine or Cobratoxin andCholinergic Receptors; 6-Aminopenicillanic acid and D-AlanineCarboxypeptidase; Plant Lectins and Epidermal Growth Factor Receptors;Alprenolol and Epinephrine Receptors; Growth Hormone and ProlactinReceptors; Insulin and Insulin Receptors; Estradiol orDiethylstilbestrol and Estrogen Receptors; Dexamethasone andGlucocorticoid Receptors; Hydroxycholecalciferol and Vitamin DReceptors. Suitable ligands include, but are not limited to, antibodies,nucleic acids, antitoxins, peptides, chelating agents, enzymeinhibitors, receptor agonists, and receptor antagonists. The term“antibody”, as used herein, means immunoglobulins such as IgA, IgG, IgM,IgD, and IgE, whether monoclonal or polyclonal in origin. The methodsfor binding and elution for the binding pairs for affinitychromatography depend on the binding pair used, and are generally wellknown in the art. As one example, solutes with polyhistidine labels maybe purified using resins including but not limited to commerciallyavailable resins such as Superflow Ni-NTA (Qiagen) or Talon CellthruCobalt (Clontech). Polyhistidine-labeled solutes may, for example, beeluted from such resins with buffers containing imidzole or glycine.Buffers for ion exchange chromatography may be selected such that thebinding pair used is soluble in the buffer. Buffers are typically singlephase, aqueous solutions, and may be polar or hydrophobic.

Resins for binding by the targeting ligand may be selected based on thetargeting ligand and the buffers to be used.

Hydrophobic Interaction Chromatography

Hydrophobic interaction chromatography (HIC) relies on hydrophobicinteractions between the solute and the stationary phase. Typically, HICis performed with buffers at high ionic strength to increase thestrength of hydrophobic interactions, and elution is achieved byreducing the ionic strength of the buffer composition, such as pH, ionicstrength, addition of chaotropic or organic agents, such as ethyleneglycol. Varying the pH of the mobile phase may also affect the chargeand thus the hydrophobicity of the substrates to effect more efficientseparation. Nonlimiting examples of resins for HIC include agarose,sepharose, cellulose, or silica particles that may be modified withbenzyl groups, linear or branched alkyl groups with any degree ofsaturation containing 2 to 50 carbon atoms, including octayl, decyl,dodecyl, tetradecyl, hexadecyl, octadecyl, and eicosyl groups. Resinscomprising hydrophobic polymers may be of particular use, as theyeliminate the need for covering the resin with hydrophobic functionalgroups. Such solid hydrophobic polymers comprise a matt of intertwinedhydrophobic polymer chains, the chains having molecular weights of fromabout 10,000 daltons to about 10,000,000 daltons. The polymer mayoptionally be porous. Suitable polymer materials include, for example,polyethylene, polypropylene, polyether sulfone, polystyrene,polydivinylbenzene, polytetrafluoroethylene, polymethyl methacrylate,polydimethyl siloxane, and blends thereof. The polymer support may be inany form, including, for example, particles, beads, cards, sheets,fibers, hollow fibers, and semipermeable membranes.

Electrochemical Measurements

Electrochemical analysis of a liquid sample typically uses electrodesthat are dipped in a liquid sample for electrochemical determination ofthe type of analyte, measurement of the analyte concentration, or both.The electrodes are spaced apart from each other, and the electrolytes inthe sample provide ionic communication between the electrodes. In amajority of situations, the sample is static during measurement; in someinstances, the sample flows through an electrochemical detector when thesample is in fluid motion, such as in the case of flow injectionanalysis. The dimensions of the electrodes may define the volume of thesample required for the measurement. The constraints relating to thevolume of the sample and the requirement of rapid measurement may callfor the use of microelectrodes, when the volume of the sample is notsufficient to cover the surface area of electrodes of conventional size.Samples that may be measured by electrochemical analysis include but arenot limited to biological fluids such as processed or unprocessed bloodor plasma, solutions of biological samples, and liquid environmentalsamples. Analytes that may be measured with electrochemical sensorsinclude, for example, blood gases (e.g. carbon dioxide, oxygen, pH,amongst others), electrolytes (e.g. sodium ions, potassium ions), andmetabolites (e.g. glucose, lactate).

Electrochemical measurements may be used to measure any reagent that canbe used in a reaction to effect electron or charge transfer to or froman electrode. Reagents include, but are not limited to, enzymes such asglucose oxidase, glucose dehydrogenase, beta-hydroxybutyratedehydrogenase, and lactate dehydrogenase; mediators such as ferrocene,ferricyanide, quinones, and the like; co-enzymes such as nicotinamideadenine dinucleotide (NAD) if necessary; ionophores; cells; smallmolecules such as glucose; or combinations of the foregoing. Thereagents typically comprise an enzyme and a mediator. A mediator is achemical species that has two or more oxidation states of distinctelectro-active potentials that allow a reversible mechanism oftransferring electrons/charge to an electrode. The enzyme reacts withthe analyte in the sample, thereby catalyzing oxidation of the analyte.The enzyme is reduced in the oxidation reaction, and the reduced enzymeis regenerated by the mediator. Alternatively, ionic species and metalions can be used in place of the enzyme to form electrochemicallydetectable compounds when they react with the analyte, such asionophores used for the ion-sensitive electrodes.

In assays where an electroactive species in a liquid sample is measuredwithout the need for any reagent at all, the conducting layerconstituting the working electrode need not have any reagent depositedthereon. As is well-known, electrochemical measurement may be carriedout by using a working electrode coupled to a reference electrode. Themeasurement can involve a change in the potential (potentiometry) or thegeneration of current (amperometry). The electrodes by themselves do notexhibit specificity to an analyte. The specificity can be imparted tothe electrode by having an enzyme (in the case of biosensor) that reactswith only one of a plurality of analytes in a mixture of analytes or byemploying a filtration technique that would selectively allow only oneof a plurality of analytes in a mixture to pass through a filtrationdevice. In electrochemical measurements of certain analytes, such asdopamine in the brain, the determination of interfering agents in a“dummy” electrode of a biosensor is one example wherein anelectrochemical measurement is carried out without the use of anyreagent on the surface of the working electrode. See, for example, U.S.Pat. No. 5,628,890, incorporated herein by reference.

In an amperometric measurement, a constant voltage is applied at theworking electrode with respect to the reference electrode, and thecurrent between the working and counter electrodes is measured. Theresponse of the electrochemical cell has two components, catalytic(glucose response component) and Faradaic (solution resistancecomponent). If the resistance of the solution is minimized, the responseof the electrochemical cell at any given time will have substantiallyhigher glucose response component, as compared with the solutionresistance component. Therefore, one is able to obtain good correlationwith the concentration of glucose from the response of theelectrochemical cell even at assay times as short as one second. If theresistance of the solution is high, the voltage experienced at theworking electrode will lag significantly from the voltage applied. Thislag is significantly higher for a two-electrode system, as compared witha three-electrode system. In the case of two-electrode system, the valueof iR between the working and the reference electrode is significantlyhigher than that in a three-electrode system. In a three-electrodesystem, no current flows between the working electrode and the referenceelectrode, and hence the voltage drop is lower. Therefore, once thecharging current (Faradaic current) decays to a minimum (within two tothree milliseconds), the current observed is all catalytic current. In atwo-electrode system, the charging current is not diminished until thevoltage at the working electrode attains a steady state (reaches theapplied voltage). Thus, in a two-electrode system, there is a slow decayof the response profile.

The passage of the electrochemical cell can be filled with a liquidsample by any of numerous methods. Filling can be carried out by, forexample, capillary attraction, chemically-aided wicking, or vacuum.Alternatively, the liquid sample can flow through the passage. Themanner of filling the electrochemical cell depends on the application,such as single use of the sensor or continuous measurements in a flowinjection analysis.

In one example, electrochemical measurements may be used to measure thelevel of glucose in a sample of blood, which can aid in determining thequantity of insulin to be administered. Glucose is typically measured byamperometrics in the presence of an enzyme that specifically usesglucose as a substrate.

An enzyme that is currently used is glucose oxydase (GOD) because it isvery specific to glucose, does not react to any other oligosaccharides,and is insensitive to temperature variations. Glucose oxydase has,however, the drawback of being very sensitive to the presence of oxygen.As a result, variations in the oxygen levels of blood samples mayprevent precise measurement of glucose levels. To reduce or eliminatethe effects of oxygen concentration, a mediator may be used toaccelerate electron transfer. Some nonlimiting examples of suchmediators include ferrocene, its derivatives, and osmium complexes, suchas those disclosed in U.S. Pat. No. 5,393,903, which is incorporatedherein by reference.

An alternate enzyme for glucose assays may be glucose dehydrogenase(GDH), which has the advantage of being insensitive to the presence ofoxygen. Glucose dehydrogenase has, however, the drawback of being lessglucose specific and of interfering with other saccharides,oligosaccharides, and oligopolysaccharides, such as maltose, whichresults in overestimation of the glucose level.

FIGS. 99 to 100 show some embodiments of electrochemical sensorconfigurations that can be adapted for use as part of probes, tips, orother components of the system for detection of analytes. Optionally,these electrochemical sensor configurations can be integrated to be partof the device. In one non-limting example, these can be part of thehardware, such as but not limited to integration with the pipette units6720 or they may be part of the cartridge or other disposable. Someembodiment may integrate these electrodes with electrochemical sensorconfiguration(s) herein to be part of a sample collection disposablewith a connector on the disposable and a matching one on the device toread signal(s), data, or information from the disposable. This linkagecan be by direct wired connection, wireless connection, or the like.

Other detection systems such as but not limited to electrochemicalsystems may allow the embodiment to work with whole blood samplesinstead of plasma. This may decrease processing time due to thegenerally much more immediate availability of whole blood sample versusplasma. This creates a consolidated protocol without as many samplepreparation steps. Optionally, electrochemical techniques may have asystem that comprises ion selective electrodes, pH type of electrodesuch as but not limited to Clark electrode, current measuring electrode,voltage measuring electrode. This may be useful for blood-gasmeasurements that may desire to engage the sample in assay measurementsoon after collection to maintain sample integrity.

Optionally, these electrodes may be integrated into the samplecollection device, into the device and the system, or only in thesystem. In one non-limiting example, the collection device may containelectrode(s) that engage the sample, the collection device may plug intothe cartridge, and the cartridge is plugged into the device.

Ion selective detectors may also be used. Ion-selective electrodes mayinteract with specific ions in a solution to generate an electricalsignal, which may be measured. Ion-selective electrodes may be used tomonitor various ions such as, fluoride, bromide, cadmium, hydrogen,sodium, silver, lead, and gases such as ammonia, carbon dioxide, oxygen,and nitrogen oxide. Ion-selective electrodes may include, for example,glass membranes, crystalline membranes, and organic polymer membranes.In some embodiments, ion-selective electrodes may be used with generalchemistry assays disclosed herein.

In some embodiments, ion selective electrodes or membranes are dopedwith certain chemicals for detecting, for example, potassium or calcium.If when using ion-selective electrodes, the final signal may be anelectrical voltage change, current change, or change in impedance.Optionally, a detector may be doped with fluorescent compounds orfluorophores such as porphyrin phosphorescence, Pd-phosphor,tris(4,7-diphenyl-1,10-phenanthroline) cation, that can detect oxygenand are sensitive to oxygen changes. In one non-limiting example, thedetector may be a PET membrane doped with fluorophores, other polymetriccompound, or western blot materials. This may be desirable where theconfirmation test uses a technique different from the initial testtechnique. It may also result in a higher integrity assay, particularlyfor time-sensitive assays. These assays may be oxygen or other gassensitive assays where there is greater risk of the loss of assayintegrity due to undesired gas exposure during assay processes that havemany steps versus those with much fewer steps.

In some embodiments, ion assays may be performed with ionophores thatare selective for certain ions. These selective ionophores may be dopedinto a substrate such as but not limited to PET. The convenience andoptionally, a lack of need for sample processing, may allow use of asample sooner after collection from a subject, and at smaller volumes.Ionophores may be used for blood analysis, including blood gas andelectrolyte profile. Some embodiments may use lateral or laminar flowstrips, such as but not limited to those similar those from Millipore,Inc., that may have membranes that are treated with ionophores toprovide the desired detection.

Multivariate Analysis

Devices and systems provided herein may be used for multivariateanalysis. This can enable the characterization of a clinical outcome ofa subject. Devices and systems provided herein may be used to aid anend-user in diagnosis, prognosis, and treatment of a clinical outcome.

Devices and systems provided herein may be used in multivariateanalysis, in some cases with the aid of a probability or referencespace. In some cases, systems and devices provided herein are configuredto collect data for use with methods provided in U.S. patent applicationSer. No. 12/412,334 to Michelson et al. (“METHODS AND SYSTEMS FORASSESSING CLINICAL OUTCOMES”), which is entirely incorporated herein byreference. In an example, the system 700 (including one or more of themodules 701-706) is configured to process samples to assist indetermining the trajectory, velocity and/or acceleration of a treatmentor the progression of a condition (e.g., health or disease condition) ofa subject. The trajectory may be indicative of the likelihood ofprogression to the clinical outcome. In another example, the system 700collects data for use in trend analysis.

All vessels (e.g., cuvettes, tips), tips, methods, systems andapparatuses described in U.S. Provisional Patent Application No.61/435,250, filed Jan. 21, 2011 (“SYSTEMS AND METHODS FOR SAMPLE USEMAXIMIZATION”), and U.S. Patent Publication No. 2009/0088336 (“MODULARPOINT-OF-CARE DEVICES, SYSTEMS, AND USES THEREOF”), are entirelyincorporated herein by reference.

EXAMPLES

The following examples are offered for illustrative purposes only, andare not intended to limit the present disclosure in any way.

Example 1: Chem 14 and Lipid Panel

A fingerstick was used to release blood from a subject. 120 microlitersof the released whole blood was collected and mixed with ananti-coagulant (EDTA or heparin—80 microliters with EDTA and 40microliters with heparin), and transferred to two separate vessels forthe two different anti-coagulant-containing samples.

Both vessels were loaded into a cartridge containing multiplefluidically isolated reagents, vessels, and tips. The cartridge wasloaded into a device provided herein containing a module containingvarious components, including a centrifuge, a pipette containingmultiple cards, a spectrophotometer, and a PMT.

Inside the device, the pipette was used to engage the EDTA-containingand heparin-containing sample vessels, and to load them into thecentrifuge. The vessels were centrifuged for 5 minutes at 1200 g, toseparate the blood cells from the blood plasma. The vessels were thenremoved from the centrifuge, and returned to the cartridge.

The pipette was used to aspirate 16 microliters of plasma from thevessel containing the EDTA-containing sample, and to deposit theaspirated EDTA plasma into an empty vessel in the cartridge. The pipettewas also used to aspirate 32 microliters of plasma from the vesselcontaining the heparin-containing sample, and to deposit the aspiratedheparin plasma into an empty vessel in the cartridge.

The pipette was used to dilute the EDTA and heparin plasma with diluentsand to yield different plasma dilutions, through step-wise and serialdilutions. For each dilution step, a diluent was first aspirated by thepipette from a vessel on the cartridge, and deposited in an emptyvessel. The sample was then added to the diluent by the pipette, and thediluent and sample were mixed. As a result of the dilution steps,ultimately, vessels containing plasma diluted 3 to 300-fold weregenerated.

The diluted plasma was used to perform all fourteen assays of a Chem 14panel [glucose, calcium, albumin, total protein, sodium (Na), potassium(K), chloride (Cl), CO2 (carbon dioxide, bicarbonate), creatinine, bloodurea nitrogen (BUN), alkaline phosphatase (ALP), alanineaminotransferase (ALT/GPT), aspartate aminotransferase (AST/GOT), totalbilirubin] and four assays of a lipid panel (LDL cholesterol, HDLcholesterol, total cholesterol, and triglycerides).

Chem 14 Panel

For the chloride assay, the pipette aspirated 10 microliters ofEDTA-containing diluted plasma, and dispensed the diluted plasma into anempty vessel. The pipette then aspirated 20 microliters of a chloridereaction mixture containing mercury nitrate, ferrous sulfate, and2,4,6-Tripyridyl-s-triazine (TPTZ) from a vessel in the cartridge, anddispensed the reaction mixture into the vessel containing the 10microliters of diluted plasma, and mixed the solutions. The assay wasincubated, and moved to the spectrophotometer, where the absorbance ofthe sample was measured at 600 nm. The measured absorbance was 2.150 ata 10 mm pathlength equivalent. This absorbance value was plotted on acalibration curve, and was determined to indicate a level of 99.0 mmol/Lchloride in the plasma sample.

For the total protein assay, the pipette aspirated 10 microliters ofEDTA-containing diluted plasma, and dispensed the diluted plasma into anempty vessel. The pipette then aspirated 20 microliters of a totalprotein assay reaction mixture containing copper (II) sulfate from avessel in the cartridge, and dispensed the reaction mixture into thevessel containing the 10 microliters of diluted plasma, and mixed thesolutions. The assay was incubated, and moved to the spectrophotometer,where the absorbance of the sample was measured at 540 nm. The measuredabsorbance was 0.089 at a 10 mm pathlength equivalent. This absorbancevalue was plotted on a calibration curve, and was determined to indicatea concentration of 5.7 g/dL total protein in the plasma sample.

For the albumin assay, the pipette aspirated 10 microliters ofEDTA-containing diluted plasma, and dispensed the diluted plasma into anempty vessel. The pipette then aspirated 20 microliters of an albuminassay reaction mixture containing Bromocresol Green from a vessel in thecartridge, and dispensed the reaction mixture into the vessel containingthe 10 microliters of diluted plasma, and mixed the solutions. The assaywas incubated, and moved to the spectrophotometer, where the absorbanceof the sample was measured at 620 nm. The measured absorbance was 0.859at a 10 mm pathlength equivalent. This absorbance value was plotted on acalibration curve, and was determined to indicate a concentration of 3.2g/dL albumin in the plasma sample.

For the aspartate aminotransferase (AST/SGOT) assay, the pipetteaspirated 15 microliters of EDTA-containing diluted plasma, anddispensed the diluted plasma into an empty vessel. The pipette thenaspirated 15 microliters of an aspartate aminotransferase assay reactionmixture containing aspartatic acid, alpha-ketoglutaric acid, malicdehydrogenase, lactate dehydrogenase, and NADH from a vessel in thecartridge, and dispensed the reaction mixture into the vessel containingthe 15 microliters of diluted plasma, and mixed the solutions. The assaywas moved to the spectrophotometer, where the absorbance of the samplewas measured at 340 nm. The assay was then incubated, and then measuredagain at 340 nm, to determine a rate of change. The rate of change wasplotted on a calibration curve, and determined to indicate aconcentration of 34.0 IU/L aspartate aminotransferase in the plasmasample.

For the potassium assay, the pipette aspirated 15 microliters ofheparin-containing diluted plasma, and dispensed the diluted plasma intoan empty vessel. The pipette then aspirated 15 microliters of apotassium assay reaction mixture containing sodium tetraphenylboratefrom a vessel in the cartridge, and dispensed the reaction mixture intothe vessel containing the 15 microliters of diluted plasma, and mixedthe solutions. The assay was incubated, and moved to thespectrophotometer, where the absorbance of the sample was measured at450 nm. The measured absorbance was −0.141. This absorbance value wasplotted on a calibration curve, and was determined to indicate aconcentration of 3.4 mmol/L potassium in the plasma sample.

For the blood urea nitrogen (BUN) assay, the pipette aspirated 10microliters of EDTA-containing diluted plasma, and dispensed the dilutedplasma into an empty vessel. The pipette then aspirated 10 microlitersof a first BUN assay reaction mixture containing urease, sodiumsalicylate, and sodium nitroprusside from a vessel in the cartridge, anddispensed the reaction mixture into the vessel containing the 10microliters of the diluted plasma, and mixed the solutions. The assaywas incubated, and then the pipette aspirated 10 microliters of a secondBUN assay reaction mixture containing sodium hypochlorite from a vesselin the cartridge, and dispensed the reaction mixture into the vesselcontaining the plasma and first BUN assay reaction mixture, and mixedthe solutions. The assay was incubated, and moved to thespectrophotometer, where the absorbance of the sample was measured at630 nm. The measured absorbance was 0.0159 at a 10 mm pathlengthequivalent. This absorbance value was plotted on a calibration curve,and was determined to indicate a concentration of 10.4 mg/dL BUN in theplasma sample.

For the bicarbonate/carbon dioxide assay, the pipette aspirated 10microliters of heparin-containing diluted plasma, and dispensed thediluted plasma into an empty vessel. The pipette then aspirated 10microliters of a first bicarbonate/carbon dioxide assay reaction mixturecontaining phosphoenolpyruvate (PEP) and phosphoenolpyruvate carboxylase(PEPC) from a vessel in the cartridge, and dispensed the reactionmixture into the vessel containing the 10 microliters of the dilutedplasma, and mixed the solutions. The assay was incubated, and then thepipette aspirated 10 microliters of a second bicarbonate/carbon dioxideassay reaction mixture containing Fast Violet B from a vessel in thecartridge, and dispensed the reaction mixture into the vessel containingthe plasma and first bicarbonate/carbon dioxide assay reaction mixture,and mixed the solutions. The assay was incubated, and moved to thespectrophotometer, where the absorbance of the sample was measured at520 nm. The measured absorbance was 3.218 at a 10 mm pathlengthequivalent. This absorbance value was plotted on a calibration curve,and was determined to indicate a concentration of 20.0 mmol/Lbicarbonate/carbon dioxide in the plasma sample.

For the glucose assay, the pipette aspirated 10 microliters ofheparin-containing diluted plasma, and dispensed the diluted plasma intoan empty vessel. The pipette then aspirated 10 microliters of a firstglucose assay reaction mixture containing glucose oxidase from a vesselin the cartridge, and dispensed the reaction mixture into the vesselcontaining the 10 microliters of the diluted plasma, and mixed thesolutions. The pipette aspirated 10 microliters of a second glucoseassay reaction mixture containing horseradish peroxidase,4-aminoantipyrine, and 4-hydroxybenzoic acid from a vessel in thecartridge, and dispensed the reaction mixture into the vessel containingthe plasma and first glucose assay reaction mixture, and mixed thesolutions. The assay was incubated, and moved to the spectrophotometer,where the absorbance of the sample was measured at 510 nm. The measuredabsorbance was 0.623 at a 10 mm pathlength equivalent. This absorbancevalue was plotted on a calibration curve, and was determined to indicatea concentration of 69.3 mg/dL glucose in the plasma sample.

For the alkaline phosphatase (ALP) assay, the pipette aspirated 10microliters of heparin-containing diluted plasma, and dispensed thediluted plasma into an empty vessel. The pipette then aspirated 20microliters of an alkaline phosphatase assay reaction mixture containingAMPPD from a vessel in the cartridge, and dispensed the reaction mixtureinto the vessel containing the 10 microliters of the diluted plasma, andmixed the solutions. The assay was incubated, and moved to thephotomultiplier tube, where the luminescence of the sample was measuredat 510 nm. The measured signal was 188,453 counts. This absorbance valuewas plotted on a calibration curve, and was determined to indicate aconcentration of 129.0 U/L alkaline phosphatase in the plasma sample.

For the calcium assay, the pipette aspirated 10 microliters ofheparin-containing diluted plasma, and dispensed the diluted plasma intoan empty vessel. The pipette then aspirated 10 microliters of a firstcalcium assay reaction mixture containing 2-amino-2-methyl-1-propanolfrom a vessel in the cartridge, and dispensed the reaction mixture intothe vessel containing the 10 microliters of the diluted plasma, andmixed the solutions. The pipette aspirated 10 microliters of a secondcalcium assay reaction mixture containing o-cresolphthalein from avessel in the cartridge, and dispensed the reaction mixture into thevessel containing the plasma and first calcium assay reaction mixture,and mixed the solutions. The assay was incubated, and moved to thespectrophotometer, where the absorbance of the sample was measured at570 nm. The measured absorbance was −0.0112. This absorbance value wasplotted on a calibration curve, and was determined to indicate aconcentration of 8.7 mg/dL calcium in the plasma sample.

For the total bilirubin assay, the pipette aspirated 10 microliters of afirst bilirubin assay reaction mixture containing sulfanilic acid from avessel in the cartridge, and dispensed the reaction mixture into anempty vessel. The pipette then aspirated 5 microliters of a secondbilirubin assay reaction mixture containing sodium nitrite from a vesselin the cartridge, and dispensed the reaction mixture into the vesselcontaining the 10 microliters of a first bilirubin assay reactionmixture, and mixed the solutions. The pipette then aspirated 15microliters of the diluted plasma, and dispensed the diluted plasma intothe vessel containing the first and second bilirubin assay reactionmixtures, and mixed the solutions. The assay was incubated, and moved tothe spectrophotometer, where the absorbance of the sample was measuredat 570 nm. The measured absorbance was 0.081 at a 10 mm pathlengthequivalent. This absorbance value was plotted on a calibration curve,and was determined to indicate a concentration of 0.8 mg/dL totalbilirubin in the plasma sample.

For the creatinine assay, the pipette aspirated 10 microliters ofEDTA-containing diluted plasma, and dispensed the diluted plasma into anempty vessel. The pipette then aspirated 10 microliters of a firstcreatinine assay reaction mixture containing glutamic dehydrogenase,alpha-ketoglutaric acid, and NADH from a vessel in the cartridge, anddispensed the reaction mixture into the vessel containing the 10microliters of the diluted plasma, mixed the solutions, and incubatedfor 5 minutes. The pipette aspirated 10 microliters of a secondcreatinine assay reaction mixture containing creatinine deiminiase froma vessel in the cartridge, and dispensed the reaction mixture into thevessel containing the plasma and first creatinine assay reactionmixture, and mixed the solutions. The assay was then moved to thespectrophotometer, where the absorbance of the sample was measured at340 nm for a set period of time, and the rate of change was determined.The rate of change was compared to a calibration curve, and wasdetermined to indicate a concentration of 1.1 mg/dL creatinine in theplasma sample.

For the sodium assay, the pipette aspirated 10 microliters ofEDTA-containing diluted plasma, and dispensed the diluted plasma into anempty vessel. The pipette then aspirated 10 microliters of a firstsodium assay reaction mixture containing lithium chloride and achelating agent from a vessel in the cartridge, and dispensed thereaction mixture into the vessel containing the 10 microliters of thediluted plasma, and mixed the solutions. The pipette aspirated 5microliters of a second sodium assay reaction mixture containingbeta-galactosidase from a vessel in the cartridge, and dispensed thereaction mixture into the vessel containing the plasma and first sodiumassay reaction mixture, and mixed the solutions. The pipette thenaspirated 5 microliters of a third sodium assay reaction mixturecontaining 2-nitrophenyl b-D-galactopyranoside from a vessel in thecartridge, and dispensed the reaction mixture into the vessel containingthe plasma and first and second sodium assay reaction mixture, and mixedthe solutions. The assay was then moved to the spectrophotometer, wherethe absorbance of the sample was measured at 570 nm and again after aset period of time, to determine a rate of change of absorbance. Therate of change was plotted on a calibration curve, and was determined toindicate a concentration of 132.0 mmol/L sodium in the plasma sample.

For the alanine aminotransferase/alanine transaminase (ALT) assay, thepipette aspirated 7.5 microliters of EDTA-containing diluted plasma, anddispensed the diluted plasma into an empty vessel. The pipette thenaspirated 7.5 microliters of a first ALT assay reaction mixturecontaining L-alanine and alpha-ketoglutaric acid from a vessel in thecartridge, and dispensed the reaction mixture into the vessel containingthe 7.5 microliters of the diluted plasma, and mixed the solutions. Themixture was incubated. The pipette then aspirated 7.5 microliters of asecond ALT assay reaction mixture containing pyruvate oxidase,4-aminoantipyrene, horseradish peroxidase, andN-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline, (ALPS) from avessel in the cartridge, and dispensed the reaction mixture into thevessel containing the plasma and first ALT assay reaction mixture, andmixed the solutions. The mixture was incubated. The pipette thenaspirated 7.5 microliters of a third ALT assay reaction mixturecontaining sodium phosphate from a vessel in the cartridge, anddispensed the reaction mixture into the vessel containing the plasma andfirst and second ALT assay reaction mixture, and mixed the solutions.The assay was then moved to the spectrophotometer, where the absorbanceof the sample was measured at 561 nm. The measured absorbance was 1.515at a 10 mm pathlength equivalent. The absorbance value was plotted on acalibration curve, and was determined to indicate a concentration of49.0 U/L ALT in the plasma sample.

Lipid Panel

For the LDL-cholesterol assay, the pipette aspirated 10 microliters ofEDTA-containing diluted plasma, and dispensed the diluted plasma into anempty vessel. The pipette then aspirated 10 microliters of a firstLDL-cholesterol assay reaction mixture containing cholesterol esterase,cholesterol oxidase, and ALPS from a vessel in the cartridge, anddispensed the reaction mixture into the vessel containing the 10microliters of the diluted plasma, and mixed the solutions. The assaywas incubated, and then the pipette aspirated 10 microliters of a secondLDL-cholesterol assay reaction mixture containing horseradish peroxidaseand 4-aminoantipyrene from a vessel in the cartridge, and dispensed thereaction mixture into the vessel containing the plasma and firstLDL-cholesterol assay reaction mixture, and mixed the solutions. Theassay was incubated, and moved to the spectrophotometer, where theabsorbance of the sample was measured at 560 nm. The measured absorbancewas 0.038 at a 10 mm pathlength equivalent. This absorbance value wasplotted on a calibration curve, and was determined to indicate aconcentration of 93.7 mg/dL LDL-cholesterol in the plasma sample.

For the HDL-cholesterol assay, the pipette aspirated 10 microliters ofEDTA-containing diluted plasma, and dispensed the diluted plasma into anempty vessel. The pipette then aspirated 10 microliters of a firstHDL-cholesterol assay reaction mixture containing dextran sulfate andALPS from a vessel in the cartridge, and dispensed the reaction mixtureinto the vessel containing the 10 microliters of the diluted plasma, andmixed the solutions. The assay was incubated, and then the pipetteaspirated 10 microliters of a second HDL-cholesterol assay reactionmixture containing cholesterol esterase, cholesterol oxidase,horseradish peroxidase and 4-aminoantipyrene from a vessel in thecartridge, and dispensed the reaction mixture into the vessel containingthe plasma and first HDL-cholesterol assay reaction mixture, and mixedthe solutions. The assay was incubated, and moved to thespectrophotometer, where the absorbance of the sample was measured at560 nm. The measured absorbance was 0.015 at a 10 mm pathlengthequivalent. This absorbance value was plotted on a calibration curve,and was determined to indicate a concentration of 40 mg/dLHDL-cholesterol in the plasma sample.

For the total cholesterol assay, the pipette aspirated 10 microliters ofEDTA-containing diluted plasma, and dispensed the diluted plasma into anempty vessel. The pipette then aspirated 20 microliters of a totalcholesterol assay reaction mixture containing cholesterol esterase,cholesterol oxidase, horseradish peroxidase, ALPS, and 4-aminoantipyrenefrom a vessel in the cartridge, and dispensed the reaction mixture intothe vessel containing the 10 microliters of the diluted plasma, andmixed the solutions. The assay was moved to the spectrophotometer, wherethe absorbance of the sample was measured at 500 nm and again after aset period of time, to determine a rate of change of absorbance. Therate of change was plotted on a calibration curve, and was determined toindicate a concentration of 120.0 mg/dL total cholesterol in the plasmasample.

For the triglycerides assay, the pipette aspirated 10 microliters ofEDTA-containing diluted plasma, and dispensed the diluted plasma into anempty vessel. The pipette then aspirated 10 microliters of a firsttriglycerides assay reaction mixture containing lipase and ALPS from avessel in the cartridge, and dispensed the reaction mixture into thevessel containing the 10 microliters of the diluted plasma, and mixedthe solutions. The pipette then aspirated 10 microliters of a secondtriglycerides assay reaction mixture containing glycerol kinase,glycerol-3-phosphate oxidase, horseradish peroxidase and4-aminoantipyrene from a vessel in the cartridge, and dispensed thereaction mixture into the vessel containing the plasma and firsttriglycerides assay reaction mixture, and mixed the solutions. The assaywas incubated, and moved to the spectrophotometer, where the absorbanceof the sample was measured at 560 nm. The measured absorbance was 0.552at a 10 mm pathlength equivalent. This absorbance value was plotted on acalibration curve, and was determined to indicate a concentration of69.2 mg/dL triglycerides in the plasma sample.

Unless otherwise noted, each of the above pipetting steps was performedusing a new pipette tip. New pipette tips were stored in the cartridge,and used pipette tips returned to the cartridge at their originallocation. Each of the incubation steps was for no more than 15 minutes.Each of the above assays was individually completed in less than 20minutes, and the total time for multiplexing all of the above assays wasless than 1 hour.

Example 2: COV of Measurements

Samples of SeraCon I (difibrinated, pooled plasma, 0.2 μm filtered;SeraCare, Inc., Milford, Mass.) containing 3, 7.9, 10.2, or 18.1 mg/dLcalcium ions were prepared. Each of the samples was separately assayedfor calcium four times on a device provided herein, following theprocedure for the calcium assay as described in Example 1 above. Aftermixing all of the reagents for each reaction and incubating thereactions, the absorbance of the reaction mixture at 570 nm was measuredin a spectrophotometer in the device. This data is provided in Table 1.

TABLE 1 Absorbance at t = 4 min Ca conc (mg/dl) Exp 1 Exp 2 Exp 3 Exp 4Avg COV (%) 3.0 0.22 0.22 0.20 0.23 0.22 5.99 7.9 0.41 0.46 0.36 0.390.41 10.08 10.2 0.51 0.52 0.48 0.49 0.50 4.15 18.1 0.74 0.70 0.63 0.770.71 8.57

As shown in Table 1, each of the different assays with each of thedifferent calcium-containing samples yielded a similar absorbance valuefor the same calcium concentration. Based on the different assays, thecoefficient of variation (COV) for the assay was determined, for each ofthe different calcium concentrations. As shown in the Table 1, for eachof the different calcium-containing samples, the COV was 10.08% orlower. In addition, the average COV for the assay was 7.20%, calculatedbased on the COV for each different calcium-containing samples(5.99+10.08+4.15+8.57/4). FIG. 102 provides a graph of absorbance at 570nm (Y-axis) vs. concentration of the calcium in the sample (X-axis),indicating a linear relationship between the values.

Example 3: Centrifuge

A centrifuge as provided herein having 4 swinging buckets and a totalcapacity of less than 500 microliters, a diameter of approximately 3inches, base plate dimensions of approximately 3.5 inches×3.5 inches,and a height of approximately 1.5 inches was loaded with 4 centrifugetubes, with 2 of them containing 60 microliters of water containing dyeand the other 2 being empty. The centrifuge was operated for 4 “highspeed” and 3 “low speed” runs, with the high speed run having a targetRPM 6.2 times greater than the low speed run. Each run was for at least180 seconds in duration. For the first 3 minutes of each centrifuge run,the RPM of the rotor was recorded every second. The coefficient ofvariation for the centrifuge was calculated. The average speed of therotor between 50 and 150 seconds for each of the high speed runs and thelow speed runs was determined Based on this data, the COV for both thehigh speed and low speed runs across the different runs was determined:COV for the low speed runs was 2.2%, and for the high speed runs was1.5%.

Example 4: Fluid Handling Apparatus

A fluid handling apparatus as provided herein having 9 pipetteblades/cards, each mounted on a common support structure, was tested forprecision and coefficient of variation of liquid transfer. 8 of the 9cards (card numbers 1-8) had the same internal configuration, optimizedfor pipetting small volumes (“low capacity” cards). 1 of the 9 cards(card number 9) had an internal configuration optimized for pipettinglarger volumes (a “high capacity” card) and had a larger internal volumeof the piston cavity and piston than the low capacity cards.

Each of the 9 cards was tested for performance of pipetting a relativelysmall volume and a relatively large volume, based on the overallcapacity of the cards. For the low capacity cards, the relatively smallvolume was approximately 2 microliters and the relatively large volumewas approximately 10 microliters; for the high capacity card, therelatively small volume was approximately 5 microliters and therelatively large volume was approximately 40 microliters. Specifically,the low capacity cards were tested for performance of aspirating anaqueous solution based on movement of the piston within the card as aresult of a first selected number of ticks and a second selected numberof ticks of an encoder wheel of the motor operatively connected to thepiston; the first selected number of ticks correspond approximately to 2microliters and the second selected number of ticks correspondapproximately to 10 microliters. The high capacity card was tested forperformance of aspirating an aqueous solution based on movement of thepiston within the card as a result of a first selected number of ticksand second selected number of ticks of an encoder wheel of the motoroperatively connected to the piston; the first selected number of tickscorrespond approximately to 5 microliters and the second selected numberof ticks correspond approximately to 40 microliters.

Performance of the each pipette cards was measured as follows. Eachpipette card aspirated an aqueous dye based on the movement of thepiston within the card as a result of the respective first or secondnumber of ticks of the motor operatively connected to the piston, asdescribed above. The dye was dispensed into known volume of water, andthe absorbance of each water-dye solution was determined. The absorbanceis directly related to the quantity of dye dispensed by the pipette cardinto the water, and it may be used to calculate the volume of dyedispensed into the water. Each pipette card performed the relativelysmall volume and the relatively large volume pipetting procedure asdescribed above 10 times, and the volume of liquid pipetted by the cardfor each procedure was determined. Then, the average volume of liquidpipetted by each card for the relatively small volume and the relativelylarge volume procedure was determined. These values are provided inTable 2. In addition, based on the variance between volumes pipetted byeach card across each of the 10 pipetting procedures for each of therelatively large volume and relatively small volume procedures, thecoefficient of variation (COV) for each pipette card for the relativelylarge volume and relatively small volume procedures was determined.These values are also provided in Table 2. As indicated in the Table,for the relatively low volume procedures, all pipette cards have a COVof 2.2% or lower. For the relatively high volume procedures, all pipettecards have a COV of 0.8% or lower. Furthermore, the average coefficientof variation between each of the 9 cards of the fluid handling apparatusfor pipetting: i) the relatively small volume and ii) the relativelylarge volume was also determined, and is provided in Table 2. As shownin the Table, across all cards in the fluid handling apparatus, theaverage COV for the relatively small volume pipetting was 1.3%, and forthe relatively large volume pipetting was 0.5%.

TABLE 2 Average Volume Coefficient of Pipetted (μl) Variation (%)Relatively Relatively Relatively Relatively Small Large Small LargeCard/Blade # Volume Volume Volume Volume 1 1.90 10.43 0.5 0.4 2 1.9310.46 1.6 0.8 3 1.92 10.40 1.7 0.5 4 1.91 10.40 0.6 0.6 5 1.90 10.41 1.00.6 6 1.93 10.41 0.9 0.6 7 1.91 10.44 1.4 0.4 8 1.91 10.39 1.7 0.6 94.80 40.56 2.2 0.4 Fluid Handling 1.3 0.5 Apparatus-Wide Average COV:

Example 5: Spectrophotometer

A spectrophotometer in a device described herein was used for variousmeasurements.

In one experiment, the spectrophotometer was used to obtain multiplemeasurements of the absorbance of different NADH-containing solutions.Solutions of 62.5, 125, 250, 500, and 1000 micromolar NADH in 20 mmTris, 0.05% sodium azide were prepared. The absorbance of each solutionat 340 nm was measured each minute for a period of 20 minutes. Theresults of the measurement at each minute for each solution are providedin FIG. 103. Based on the 20 measurements for each different NADHsolution, a COV of variation for measurement of each of the solutionswas determined, and is provided below in Table 3. Table 3 also providesthe average absorbance measurement and the standard deviation for eachsolution.

TABLE 3 NADH, uM Average A340 nm St. Dev % CV 62.5 0.0622 0.0037 5.88125 0.1531 0.0039 2.56 250 0.3390 0.0043 1.26 500 0.7170 0.0053 0.741000 1.2524 0.0075 0.60

In another experiment, solutions of 62.5, 125, 250, 500, and 1000micromolar NADH in 10 mm potassium phosphate, pH 8.0 were prepared. Theabsorbance of each of the solutions at 340 nm was measured in both: i) aSPECTROstar Nano (BMG Labtech) plate reader (“commercial plate reader”);and ii) a spectrophotometer provided herein (“Theranos”) in a deviceprovided herein. The absorbance measurements by each device was plotted(X-axis: concentration of NADH in micromolarity; Y-axis: absorbance ofthe sample at 340 nm), and the slope for the values from each device wascalculated (FIG. 104). For the SPECTROstar Nano, the calculated slopewas: y=0.0018x+0.0289; R²=0.9937. For the Theranos spectrophotometer,the calculated slope was: y=0.0013x+0.0129; R²=0.9934. The high R² valuefor slope based on values from the Theranos spectrophotometer indicatesthe linearity of the spectrophotometer for measuring absorbance across awide range of concentrations of solution.

In another experiment, samples containing different amounts of urea wereassayed for absorbance. Samples of SeraCon I (difibrinated, pooledplasma, 0.2 μm filtered; SeraCare, Inc., Milford, Mass.) containingapproximately 1, 16, 23, or 71 milligrams/deciliter blood urea nitrogenwere assayed for urea. The absorbance of each of the assays at 630 nmwas measured in both: i) a SPECTROstar Nano plate reader (“commercialplate reader”); and ii) a spectrophotometer provided herein (“Theranos”)in a device provided herein. The absorbance measurements by each devicewas plotted (X-axis: concentration of blood urea nitrogen in sample inmg/dl; Y-axis: absorbance of the sample at 630 nm), and the slope forthe values from each device was calculated (FIG. 105). For theSPECTROstar Nano, the calculated slope was: y=0.0058x+0.0854; R²=0.9985.For the Theranos spectrophotometer, the calculated slope was:y=0.0061x+0.0358; R²=0.9974. The high R² value for slope based on valuesfrom the Theranos spectrophotometer indicates the linearity of thespectrophotometer for measuring absorbance across a wide range ofconcentrations of solution.

Referring now to FIGS. 106 and 107, one embodiment of a module 10100suitable for use in a rack or other common mounting structure will nowbe described. FIG. 106 is a top-down view of some components in a module10100. In this non-limiting example, a gantry system 10102 that providesX-Y axis or other axis movement of a pipette 10104 is shown in phantom.The pipette 10104 may be one such as that shown in FIGS. 66 to 67D. Thegantry system 10102 may move as indicated by arrow 10106. In oneembodiment, pipette 10104 can move as indicated by arrow 10108. Thiscombination of the gantry 10102 and pipette 10104 allows for movement inat least the XYZ axis, allowing for the movement of sample vessels toand from multiple locations in the module. FIG. 106 also shows that thepipette 10104 in a second location, or optionally, some systems may usesecond pipette and gantry system with the module 10100.

FIG. 106 shows that there may be an assay station receiving location10110 configured to receive a cartridge. In one non-limiting example,the assay station receiving location 10110 may be a tray that is movableas indicated by arrow 10112 by the use of motor 10114 and gear tracks10116 to move the tray outside of the module to facilitate userplacement of one or more cartridges into the module.

Once a cartridge is in the system, individual elements of the cartridgesuch as but not limited to cuvettes, pipette tips, vessels, otherphysical items, regent(s), fluids, or the like may be moved from thecartridge. FIG. 106 also shows that there may be a variety of componentsin the module 10100 such as but not limited to a centrifuge 10120, ahigh sensitivity optical detector 10122 such as but not limited to aPMT, a multi-array optical detector 10124 such as but not limited to aspectrophotometer, and a nucleic acid amplification module 10126. Eachof these components may have its own sample vessel receiving locationsuch as but not limited to locations 10130, 10132, 10134, and 10136. Inone non-limiting example, the locations 10130, 10132, 10134, and 10136may be sized to be different shapes, sized to receive different types ofvessels, and in the case of the centrifuge, may have a variable locationdepending on where the centrifuge finishes spinning A controller of thesystem is configured to direct sample vessels to the desired locationsand be able to accurately place them in the appropriate receivinglocations for each of the components.

FIG. 107 shows a side view of the various components in the module10100.

It should also be understood that thermal control of conditions withinthe module 10100 can be regulated so that thermal conditioning by way ofcontrolled temperature air flow through the system is accomplished sothat temperature sensor(s) in the module detect that ambient air in thesystem is within a desired range. Optionally, the thermal regulation isby way of a combination of controlled air temperature and controlledsupport structure temperature. This can be of particular use when thesupport structure comprises of a thermally conductive material.

Referring now to FIG. 108, one embodiment of a system 10200 with linkedconvective flow between modules will now be described. As seen in FIG.108, the common flow between modules 10100 is shown by arrow 10202.Inlet air flow into each of the modules 10100 is indicated by arrow10204. Optionally, the thermal conditioning of adjacent modules can beused to condition the underside or other surfaces of adjacent modules.In this manner, combined module thermal conditioning can create a morestable thermal state for all of the modules sharing a common mounting.This convective air flow within a module is indicated by arrow 10208.Optionally, a convective flow unit 10220 which may provide thermallyconditioned (heated, cooled, or neutral) airflow can be used to maintaina desired air temperature range within the substantially light tightconfines of the modules 10100. One more temperature sensors 10230 may beincluded in the modules 10100 to provide feedback to a controller toadjust flow rate and/or air temperature coming from device 10220. Afully or at least partially enclosed pathway 10232 may be used to directexhaust air flow to a filtered outlet 10234 that may have an exhaust fantherein. Optionally, flow can be reversed on the exhaust fan such thatit can also function as an inlet if the fan is operated in reversed.

Referring now to FIGS. 109 and 110, optionally, some embodiments mayhave a bilayer module configuration wherein certain hardware elementsare mounted on a first plane while second elements that may have adifferent height are mounted on a second plane such that the features onthe first plane and second plane have sample vessel loading areas inzones or planes accessible by a common pipette system mounted on an XYgantry. By way of non-limiting example, Figure AH1 shows that an opticaldetector component 10250 may have an upper surface 10252 that is locatedwithin the range of motion of the sample handling system with gantry10102. In one non-limiting example, the upper surface 10252 is above thefirst support layer 10260 while the device 10250 is mounted on thesecond support layer 10262. The surface 10252 may be sized to receiveone cuvette or multiple cuvettes. FIG. 109 also shows that the pipette10104 in a second location, or optionally, some systems may use secondpipette and gantry system with the module 10100. It should be understoodthat the housing 10270 may be a light-tight housing. Some embodimentsmay align a plurality of the bilayer modules in a stack (similar to FIG.108) and/or horizontal combination wherein all of the resources arecontained in each of the bilayer modules. Some may not use anyadditional transport devices between bilayer modules, but such transportdevices are not excluded in alternative embodiments.

FIGS. 106 to 110 show non-limiting examples of configurations of modulesaccording to embodiments described herein.

The primary challenge in being able to accurately measure blood gasconcentrations is in maintaining the integrity of sample starting fromcollection and through sample processing, reaction, and signal read. Thegoal is to minimize mass transfer of blood gas components to and fromthe sample. In what follows, the different steps where the sample haspotential to come in contact with air are detailed, along with ways tominimize or eliminate mass transfer. Although this example is discussedin the context of measuring blood gas, it should be understood that thisis also applicable to other assays where the combination of structure inthe hardware, structure in the disposable, processing techniques in thesystem, and specific chemistries in the vessels can be combined in oneor more sequences to perform assays not otherwise possible intraditional settings due to the lack of system integration and variableapplication of the combinations of factors and capabilities herein.

Penetrating the sealed sample vessel and depositing into the centrifugevessel or other sample vessel. Optionally, the system skips the sealedsample vessel and deposits arterial blood from a syringe into thecentrifuge vessel or sample vessel. Optionally, the centrifuge vessel orsample vessel has a re-sealable seal, septa, or other seal on it tomaintain a gas tight environment therein. The seal may be polypropylene,foil polypro combination, rubber, or any material that can reseal afterbeing penetrated. When the needle tip penetrates the seal, the seal orsepta on the centrifuge vessel or sample vessel can maintain theenvironment therein without loss of integrity of the atmosphere therein.If the pressure difference between the sample (atmospheric pressure),and the pressure inside the sample vessel is high or the speed at whichfluid is transferred is high, this results in turbulent mixing in thefluid. This can increase mass transfer between the sample and air. Inone embodiment, lowering the pressure drop will result in a more gradualfill, which minimizes mixing.

In some embodiments, particularly those not collected from an arterialsample into a syringe, the sample collected in the vessel is in contactwith air which fills up the rest of the sample vessel. This can resultin mass transfer between sample and air. One method to circumvent thisis to pre-fill the sample with an inert liquid which is immiscible with,and has a lower density than the sample. By doing so, when the sample iscollected, it displaces the less dense liquid to the top, therebyforming a liquid barrier between the sample and air. There are severaloptions for this liquid barrier. Simple examples include alkane solventssuch as hexane, heptane, decane, and cyclohexane. Optionally, some mayuse fluorohydrocarbon materials. The choice of barrier fluid is mostlybased on chemical compatibility with the sample. In addition, low oxygensolubility of the barrier fluid is preferred to further reduce anypossibility of mass transfer. An example of a low-oxygen solubilitybarrier is EPDM liquid copolymer (ethylene propylene diene monomer).Long-chain fatty acid based surfactants and proteins (eg. Whey protein)also act like liquid barriers. Optionally, oxygen scavengers embeddedinto the polymer matrix is an option, such as used in the food packagingindustry. Optionally, a transition metal (iron, Cobalt, Nickel etc.)embedded into a polymer such as PET, PP, HDPE along with an activatingcomponent (electrolytes such as NaCl, electrolytic acidifying component,Na Bisulfate) can promote the reaction of the oxidizable metal with O2.Any single or multiple combination of the foregoing may be used.

Then the vessel may go to a sample separation device such as but notlimited to a centrifuge or magnetic separation facility, which performsthe separation. The vessel is then returned to an assay station such asbut not limited to a cartridge. The seal or septum on the sample vesselis then penetrated again by a liquid head pipette needle or tip,extracts the sample, deposits the sample in an detector vessel such asbut not limited to a colorimetry vessel, cuvette, or clinical chemistryvessel. This vessel is also sealed from the external atmosphere. Inanother configuration, this vessel is not sealed. In the sealedconfiguration, it has in it a set of reagents that are oxygen depletedand pre-mixed where they have already been oxygen depleted and sealed inthis vessel or cuvette. Then, the seal can be resealable or not,depending on the embodiment. This seal is punctured by a liquid headpipette needle or tip that comes in. The sample is deposited in thevessel with the oxygen depleted reagents and begins reacting. In an openvessel, some chromogen on the top portion of the mixture beginsabsorbing oxygen, but the oxygen reading is occurring in the lowerportion of the sample and is not impacted by the upper surfaceinteraction with oxygen. Optionally, in a seal vessel, this chromogeninteraction with outside air is less of an issue, particularly if sampleis still being read from the bottom of the vessel.

Starting from separation (such as through centrifugation), plasmaextraction, dilution, mixing with reagents, incubation of reactionmixture, and finally signal read. Mass transfer can be minimized in allthese steps by utilizing the same barrier fluid in all vessels thesample is transferred into. This is especially critical in the first fewstages of centrifugation, plasma extraction, and dilution, where neatplasma is handled. Having a liquid barrier prevents the sample fromcoming in direct contact with air. The liquid barrier also allows forroutine pipette operations such as extraction and mixing to be performedas normal.

Optionally, the cartridge may have a tip that has a hook, such as butnot limited to a harpoon shaped tip, that can pierce through the seal atthe top of the vessel and remove the entire plug or seal so that theentire vessel may be more easily accessible for sample removal using alarger volume tip that will less agitate the sample as it is beingtransferred. Optionally, some may have a needle tips to pierce throughthe seal on the sample vessel without removing the cap or seal.

Some embodiments may have a knife or cutting attachment that may beengaged by a pipette nozzle. This can be of particular use whenpreparing tissues for slides or staining. The pipette or otherend-effector in the system can use one nozzle with a cutting tip to cutwhile one or more other nozzles can engage the tissue directly orthrough a tip, cuvette, tissue holder, or the like to cut the tissue.

For body fat measurement, it should be understood that some embodimentsmay use not just the touch screen. Some may have other locations on thesystem for the user to contact such as an electrode or the like.

It should be understood that some cartridges with only a single rail canalso be engaged to the cartridge receiving location in a manner so thatthe cartridge can read the materials therein. Pushing the cartridgealong one rail until it reaches an alignment location registers thelocation of the cartridge in a manner that the system can then processbased on machine vision or system configuration of the cartridge IDallows the system to determine the type and configuration of thecartridge that is inserted. This correlation may be based on informationon board the device or based on information retrieved based on lookup ona remote server.

The publications discussed or cited herein are provided solely for theirdisclosure prior to the filing date of the present application. Nothingherein is to be construed as an admission that the present invention isnot entitled to antedate such publication by virtue of prior invention.Further, the dates of publication provided may be different from theactual publication dates which may need to be independently confirmed.All publications mentioned herein are incorporated herein by referenceto disclose and describe the structures and/or methods in connectionwith which the publications are cited. The following applications arealso incorporated herein by reference for all purposes: U.S. Pat. Nos.7,888,125, 8,007,999, 8,088,593 and U.S. Publication No., US20120309636,PCT Application No. PCT US2012/057155, U.S. patent application Ser. No.13/244,952, and PCT Application No. PCT/US2011/53188, filed Sep. 25,2011. PCT Application No. PCT/US2011/53188, filed Sep. 25, 2011, U.S.patent application Ser. No. 13/244,946, filed Sep. 26, 2011, PCTApplication No. PCT/US11/53189, filed Sep. 25, 2011, Patent CooperationTreaty Application No. PCT/US2011/53188; Patent Cooperation TreatyApplication No. PCT/US2012/57155; U.S. patent application Ser. No.13/244,947; U.S. patent application Ser. No. 13/244,949; U.S. patentapplication Ser. No. 13/244,950; U.S. patent application Ser. No.13/244,951; U.S. patent application Ser. No. 13/244,952; U.S. patentapplication Ser. No. 13/244,953; U.S. patent application Ser. No.13/244,954; U.S. patent application Ser. No. 13/244,956; and U.S. patentapplication Ser. No. 13/769,779, entitled “Systems and Methods forMulti-Purpose Analysis,” filed Feb. 18, 2013, all of which applicationsare hereby incorporated by reference in their entireties.

While preferred embodiments of the present invention have been shown anddescribed herein, it will be obvious to those skilled in the art thatsuch embodiments are provided by way of example only. Numerousvariations, changes, and substitutions will now occur to those skilledin the art without departing from the invention. It should be understoodthat various alternatives to the embodiments of the invention describedherein may be employed in practicing the invention. Any feature, whetherpreferred or not, may be combined with any other feature, whetherpreferred or not. The appended claims are not to be interpreted asincluding means-plus-function limitations, unless such a limitation isexplicitly recited in a given claim using the phrase “means for.” Itshould be understood that as used in the description herein andthroughout the claims that follow, the meaning of “a,” “an,” and “the”includes plural reference unless the context clearly dictates otherwise.For example, a reference to “an assay” may refer to a single assay ormultiple assays. Also, as used in the description herein and throughoutthe claims that follow, the meaning of “in” includes “in” and “on”unless the context clearly dictates otherwise. Finally, as used in thedescription herein and throughout the claims that follow, the meaning of“or” includes both the conjunctive and disjunctive unless the contextexpressly dictates otherwise. Thus, the term “or” includes “and/or”unless the context expressly dictates otherwise.

What is claimed is:
 1. A point-of-service, benchtop biological sampleprocessing device comprising: a housing containing therein: an automatedsample handling system comprising a fluid handling apparatus comprisinga pipette having a nozzle configured to: a) engage and hold a pipettetip and b) to engage and hold a microscopy cuvette, wherein saidmicroscopy cuvette is configured to engage said pipette nozzle; a firstdetection station comprising an electronic optical sensor; a seconddetection station comprising a light source and an electronic opticalsensor or sensing device; a cytometry station comprising an imagingdevice and a stage for receiving said microscopy cuvette; a centrifuge;a convective flow unit to provide thermally conditioned airflow tomaintain a desired air temperature range in the housing; one or moretemperature sensors to provide feedback to a controller to adjust airtemperature; a fan for exhausting air flow through a filtered outlet onthe housing; a cartridge receiving location; wherein the automatedsample handling system is configured to have fluid handling apparatusmovable to individually access each of the following: first detectionstation, second detection station, cytometry station, centrifuge, andcartridge receiving location.
 2. The biological sample processing deviceof claim 1, wherein the biological sample has a volume of no greaterthan 500 microliters.
 3. The biological sample processing device ofclaim 1, wherein the second detection station comprises a nucleic acidassay station, wherein the light source and electronic optical sensor orsensing device of the second detection station are contained within thenucleic acid assay station.
 4. The biological sample processing deviceof claim 3, wherein the nucleic acid assay station comprises a thermalblock.
 5. The biological sample processing device of claim 3, whereinthe nucleic acid assay station comprises at least a first well and asecond well, and the first well is configured for the measurement of thefluorescence of samples and the second well is configured for themeasurement of the turbidity of samples.
 6. The biological sampleprocessing device of claim 1, wherein the biological sample processingdevice further comprises a third detection station, wherein thecartridge further contains a fourth assay unit, wherein the fourth assayunit comprises gear teeth, and wherein the biological sample processingdevice is configured to transport the fourth assay unit to the thirddetection station by using a gear mechanism to engage the gear teeth inthe assay unit.
 7. The biological sample processing device of claim 1,wherein the biological sample processing device further contains thereina barometer.
 8. The biological sample processing device of claim 1,wherein the electronic optical sensor is selected from the group ofelectronic optical sensors consisting of photodiodes, photomultipliertubes (PMTs), Charge-Coupled Devices (CCDs), super-cooled CCD arrays,CMOS sensors, photodetectors, photon counting detectors, avalanchephotodiodes, avalanche photodiode arrays, pin diodes,spectrophotometers, and electronic cameras.
 9. A method for performingmultiple assays with a biological sample, the method comprising:providing a point-of-service, benchtop biological sample processingdevice comprising a housing containing therein: an automated samplehandling system comprising a fluid handling apparatus comprising apipette having a nozzle configured to: a) engage and hold a pipette tipand b) to engage and hold a microscopy cuvette, wherein said microscopycuvette is configured to engage said pipette nozzle; a first detectionstation comprising an electronic optical sensor; a second detectionstation comprising a light source and an electronic optical sensor; acytometry station comprising an imaging device and a stage for receivingsaid microscopy cuvette; and a cartridge, wherein the cartridge isinsertable into and removable from the biological sample processingdevice, and wherein the cartridge contains i) a biological sample, ii)at least a first, a second, and a third fluidically isolated assay unit,and iii) all reagents that are required to perform a first, second, andthird assay comprising A) at least one luminescence assay; B) at leastone absorbance, turbidimetric, fluorescence, or colorimetric assay; andC) at least one cytometry assay, and wherein the first assay unit isconfigured to perform a luminescence assay, the second assay unit isconfigured to perform an absorbance, turbidimetric, fluorescence, orcolorimetric assay, the third assay unit is configured to perform acytometry assay, and transferring, by the fluid handling apparatus ofthe automated sample handling system, said microscopy cuvette to saidstage of said cytometry station; transferring by the automated samplehandling system at least a portion of the biological sample to each ofthe first, second, and third assay units; transferring by the automatedsample handling system the first assay unit containing biological sampleto the first detection station; transferring by the automated samplehandling system the second assay unit containing biological sample tothe second detection station; transferring by the automated samplehandling system the third assay unit containing biological sample to thecytometry station, wherein said third assay unit comprises saidmicroscopy cuvette configured to perform a cytometry assay, saidmicroscopy cuvette having at least one cavity configured to accept asample, and having at least one pick-up interface configured to engagewith said fluid handling apparatus, said transferring comprising saidpipette nozzle engaging and holding said pick-up interface of saidmicroscopy cuvette and moving said microscopy cuvette by said pipettewhile said microscopy cuvette is held by said pipette nozzle; andtransferring from the cartridge to the first, second, and third assayunits, respectively, all reagents required for the performance of thefirst, second, and third assays to the respective assay units; whereinthe sample processing device also contains a centrifuge; a convectiveflow unit to provide thermally conditioned airflow to maintain a desiredair temperature range in the housing; one or more temperature sensors toprovide feedback to a controller to adjust air temperature; a fan forexhausting air flow through a filtered outlet on the housing.
 10. Themethod of claim 9, wherein the biological sample has a volume of nogreater than 500 microliters.
 11. The method of claim 9, wherein aturbidimetric assay is performed in the second assay unit, and whereinthe turbidity of the turbidimetric assay is measured over a period oftime.
 12. The method of claim 9, wherein the electronic optical sensoris selected from the group of electronic optical sensors consisting ofphotodiodes, photomultiplier tubes (PMTs), Charge-Coupled Devices(CCDs), super-cooled CCD arrays, CMOS sensors, photodetectors, photoncounting detectors, avalanche photodiodes, avalanche photodiode arrays,pin diodes, spectrophotometers, and electronic cameras.
 13. The methodof claim 9, wherein the cytometry assay comprises determining the numberof at least one type of cell per selected volume of biological sample,the at least one type of cell being selected from the group consistingof: monocytes, lymphocytes, neutrophils, basophils, and eosinophils. 14.The method of claim 9, wherein the biological sample is from a subject,and wherein prior to insertion of the cartridge into the biologicalsample processing device, the subject's insurance coverage is checked.15. The method of claim 9, wherein the biological sample processingdevice further comprises a third detection station, wherein thecartridge further contains a fourth assay unit, wherein the fourth assayunit comprises gear teeth, and wherein the method further comprisestransporting the fourth assay unit to the third detection station byrotation of a gear mechanism, wherein rotation of the gear mechanismengages the gear teeth in the fourth assay unit and moves the fourthassay unit into the third detection station.
 16. The method of claim 9,wherein the cartridge comprises a lid and a locking feature, wherein thelid comprises an engagement feature which is configured to optionallyengage the locking feature in the cartridge, and wherein the engagementfeature does not engage the locking feature in the cartridge uponinsertion of the cartridge into the sample processing device.
 17. Themethod of claim 9, wherein the second detection station comprises anucleic acid assay station, wherein the light source and electronicoptical sensor or sensing device of the second detection station arecontained within the nucleic acid assay station.